CA2480404A1 - Fc receptor homolog, reagents, and uses thereof - Google Patents

Abstract

The invention relates to members of the Fc receptor homolog (FcRH) subfamily, as well as fragments and variants thereof. Each FcRH is a Type I transmembrane receptor, preferably, comprises an extracellular region, a transmembrane region, and a cytoplasmic region. The cytoplasmic region preferably comprises one or more immunoreceptor tyrosine-based inhibitory or activation motifs ("ITIMs" or "ITAMs). The invention provides polypeptides, nucleic acids, vectors, expression systems, and antibodies and antibody fragments related to the FcRHs as well as uses thereof. Such uses include uses in the diagnosis and treatment of a malignancy of hematopoietic cell lineage or an inflammatory or autoimmune disease in a subject and in the modulation of a humoral immune response in a subject.

Description

MEMBERS OF THE FC RECEPTOR HOMOLOG GENE FAMILY
(FCRHl-3, 6), RELATED REAGENTS, AND USES THEREOF
This application claims the benefit of U.S. Provisional Application No.
60/367,667, filed March 25, 2002.
ACKNOWLEDGEMENTS
This invention was made with government support under Grants 2837 and AI39816 awarded by NIAID. The government has certain rights in the invention.
FIELD OF THE INVENTION
This invention relates generally to immunology and modulation of immunologic responses in the context of inflammatory diseases and cancer.
BACKGROUND OF THE INVENTION
Receptors for the Fc region (FcRs) of Igs have broad tissue distribution patterns and can modulate cellular and humoral immunity by linking their antibody ligands with effector cells of the immune system (Ravetch, J. V. & Kinet, J.-P. (1991) Annu. Rev.
Immunol. 9, 457-492; Daeron, M. (1997) Annu. Rev. Immunol. 15, 203-234. These cellular, receptors have the ability to sense humoral concentrations of antibody, initiate cellular responses in host defense, and participate in autoimmune disorders (Ravetch, J.
V. & Bolland, S. (2001) Annu. Rev. Immunol. 19, 275-290). Their diverse regulatory roles depend on the Ig isotype specificity and cellular distribution of the individual FcR.
These Ig superfamily members share similarities in their ligand binding subunits, and they may have inhibitory or activating signaling motifs in their intracellular domains or instead pair with signal transducing subunits possessing activating signaling motifs.
Recently, characterization of FcR hornologs in mice, the paired Ig-like receptors (Kubagawa, H. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 5266; Hayami, K. et al. (1997) J. Biol. Chem. 272, 7320-7327), and their relatives in humans the Ig-like transcripts/ leucocyte Ig-like receptors (Borges, L. et al.
(1997) J.
Immunol. 159, 5192-5196; Samaridis, J. & Colonna, M. (1997) Eur. J. Immunol.
27, 660-665) have been elucidated. This multigene family, which includes the FcaR
(Kremer, E. J. et al. (1992) Hum. Genet. 89, 107-108) and the natural killer cell Ig-like receptors (Wagtmann, N. et al. (1997) Curr. Biol. 7, 615-618), is located in a human chromosome 19q13 region lcnown as the leucocyte receptor complex (LRC) (Wende, H.
et al. (1999) Mamm. Genome 10, 154-160; Wilson, M. J. et al. (2000) Proc.
Natl. Acad.
Sci. USA 97, 4778-4783). These Ig-like multigene families belong to a larger class of receptors characterized by their possession of common cytoplasmic tyrosine-based signaling motifs. These can be either immunoreceptor tyrosine-based activation motifs (ITAMs) containing two repeats of the consensus sequence Y-X-X-L/I spaced by 6-amino acids (E/D)-X-X-Y-X-X-(L/I)-X~_8-Y-X-X-(L/IJ (SEQ ID N0:64, with six amino acid between the consensus sequences; SEQ ff~ N0:65, with seven amino acid residues between the consensus sequences; and SEQ ID N0:66, with eight amino acid residues between the consensus sequences) or immunoreceptor tyrosine-based inhibitory motifs (ITIMs) with a 6-amino acid consensus sequence (I/V/L/S)-X-Y-X-X-(L/V) (SEQ ID
N0:67) (Beth, M. (1992) Annu. Rev. Immunol. 10, 97-121; Vely, F. & Vivier, E.
(1997) J. Immunol. 159, 2075-2077; Ravetch, J. V. ~ Lamer, L. L. (2000) Science 290, 84-89; Gergely, J. et al. (1999) Immunol. Lett. 68, 3-15). The phylogenetic conservation of these types of receptors in birds (Dennis, G. et al. (2000) Proc. Natl.
Acad. Sci. USA
97, 13245-13250) and bony fish (Yoder, J. A. et al. (2001) Proc. Natl. Acad.
Sci. USA
98, 6771-6717) is indicative of their biological value. After ligand binding of the activating receptor complexes, TTAM tyrosines are rapidly phosphorylated by Src family kinases to initiate a cascade of signaling events that trigger cellular activation. In the case of ITIM-bearing receptors, the tyrosines provide a docking site for phosphatases containing Src homology 2 domains that can abrogate cellular activation (Long, E. O. (1999) Annu. Rev. Immunol. 17, 875-904; Unkeless, J. C. & Jin, J.
(1997) Curr. Opin. hnmunol. 9, 338-343). The balance in the utilization of these activating and inhibitory receptor pairs can serve to modulate cellular responses to a variety of stimuli.
The genes encoding the classical FcyRs, FcyRI, Fc~yRII, FcyRIII, and FcsRI, lie on the long arm of chromosome 1 (1q21-23) near the polymeric Ig receptor (pIgR) and Fca/~,R genes (1q32) (20-23). Members of this FcR subfamily have relatively low extracellular homology with the FcR-related genes that reside in the LRC on chromosome 19. Like the FcyR- and FcsR-activating receptors, the ligand binding chain of the FcaR coassociates with the ITAM containing FcR common y-chain (Pfefferkorn, L. C. & Yeaman, G. R. (1994) J. Immunol. 153, 3228-3236; Morton, E.
C. et al. (1995) J. Biol. Chem. 270, 29781-29787). New members of the FcR
family were sought which could have diverse signally properties and oncogenic potential.
SUMMARY OF THE INVENTION
In accordance with the purposes) of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to members of a cluster of FcR
and FcR gene relatives encoded, for example, by genes in the human chromosome 1q21-23 region, or analogous region in non-human subjects. The members are Type I
transmembrane receptors, or alternatively spliced forms thereof, with homology to the FcR family and are referred to herein as FcRHs. Each FcRH can comprise an extracellular region, a transmembrane region, and a cytoplasmic region. The cytoplasmic region preferably comprises one or more immunoreceptor tyrosine-based inhibitory or activation motifs ("ITIMs" or "ITAMs).
The invention relates to polypeptides corresponding to isolated FcRHs (e.g., huFcRH 1, 2, 3, and 6 and moFcRHl, 2, and 3), as well as fragments and isoforms thereof. The invention further relates to nucleic acids that encode the FcRHs, as well as hybridization probes related thereto and complementary sequences. The invention further provides vectors and cells related to the nucleic acids of the invention.
The invention further relates to making an FcRH, or a fragment or variant thereof, comprising culturing a cell comprising a vector of the invention under conditions permitting expression of the FcRH. The invention also provides an antibody reagent kit comprising the antibody, or a fragment or variant thereof, and reagents for detecting binding of the antibody, fragment, or antibody variant to a ligand.
The invention further relates to uses of the polypeptides, nucleic acids and antibodies of the invention. For example, the invention relates to methods of diagnosing and methods of treating a malignancy of hematopoietic cell lineage or an inflammatory or autoimmune disease in a subject. The invention also relates to modulation of a humoral immune response in a subject.
Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
BRIEF DESCRIPTION OF THE DRAWINGS
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate (one) several embodiments) of the invention and together with the description, serve to explain the principles of the invention.
Figure 1 shows the relative position of the FcRH locus within the FcR cluster on chromosome 1. The cytogenetic location of the FcR genes is approximated from the GenBank Mapview database. The BAC clones (4, GenBank accession no. AL139409;

3, GenBank accession no. AL356276; 2, GenBank accession no. AL135929; and 1, GenBank accession no. AL353721) that span the locus are oriented in relation to their respective FcRH genes (shaded area).
Figure 2 shows the structural and sequence diversity of FcRHl, FcRH2, and FcRH3. Figure 2A is a schematic representation of FcRH molecules. The three cDNAs encode type I transmembrane proteins with similar extracellular domains, but different cytoplasmic regions. The extracellular (EC) regions contain different numbers of C2-like Ig domains and potential sites of N-linked glycosylation. The transmembrane (TM) domains are uncharged, except in the case of FcRHl. The cytoplasmic (CY) region of FcRHl contains two ITAMs (light gray boxes) and one ITAM-like region (small, lined box), whereas FcRH2 contains one ITAM and two ITIMs (dark gray boxes). FcRH3 has a long cytoplasmic tail with one ITAM, one IT1M, and an ITAM-like region. The amino acid length of each region is indicated. Figure 2B shows the multiple alignment comparison of FcRHl, FcRH2, and FcRH3 amino acid sequences (one-letter code) based on the FcRH3 sequence. Amino acid identity is represented by dots, and gaps are indicated by dashes. Predicted N-linked glycosylation sites and transmembrane domains are underlined in black. Consensus ITAM (bold) and ITIM (bold, underlined) motifs are indicated. Putative structural domains are labeled: SP, signal peptide; EC, extracellular domain; MP-TM, membrane proximal-transmembrane; and CY, cytoplasmic regions.
Amino acid lengths are indicated in parentheses.
Figure 3 shows a composite analysis of the extracellular homology among FcRH and FcR family members. Pairwise analysis of individual Ig-like subunits was performed with the CLUSTAL method algorithm using FcRH3 as the index of comparison. Individual homologous domains are coded to indicate relatedness.
Percent amino acid identities for related domains are indicated and aligned in relation to the comparative FcRH3 subunit. The amino acid identity for the membrane proximal domains (light gray subunits) of FcRHS are provided as the range of identity for all individually related domains. Comparisons that are not applicable are left blank. Amino acid sequences were derived from IRTA1 (GenBank accession no. AF343659), IRTA2 (GenBank accession no. AF34364), moFcRH (GenBank accession no. AAG28775) FcyRI (GenBank accession no. AAA35678), FcyRII (Swiss-Prot accession no.
P31994), Fc~yRIII (Swiss-Prot accession no. P08637), FcsRI (Swiss-Prot accession no.
P12319), and FcaRI (Swiss-Prot accession no. P24071).
Figure 4 shows the relative location of the mouse FcR family. Location is indicated in reference to the human FcR related genes at Ch 1q21-23 and their orthologous loci on mouse Ch 3 and Ch 1. The microsatellite marker d3Mitl87 is located within moFcRHl.
Figure 5 shows the multiple alignment comparisons of huFcRHI-5 and mouse FcRHl and 2 amino acid sequences (one-letter code) based on the FcRH3 sequence.
Amino acid gaps are indicated by dashes. Consensus ITAM (underlined) and ITIM
(italic, underlined) motifs are indicated. Amino acid lengths are indicated in parentheses.
Figure 6 shows domains marked to indicate relatedness of the Ig-like subunits.
Ig-like domain homology was determined by generation of a phylogenetic tree using DNAStar software with the CLUSTAL program and assigning arbitrary colors to individual Ig-domains of a given branch. Amino acid identities for full length, extracellular and, cytoplasmic domain comparisons are based on huFcRH3.
Closest cytoplasmic relatives are indicated in parentheses. Most identical extracellular comparisons between mouse and human relatives are highlighted in horizontal lines.
Comparisons that are not applicable are left blank.
Figure 7 shows the domains of huFcRHI-6, moFcRHl-3 and related proteins.
Domains are colored to indicate relatedness of the Ig-like subunits. Ig-like domain homology was determined by generation of a phylogenetic tree using DNAStar software with the CLUSTAL program and assigning arbitrary colors to individual Ig-domains of a given branch. Amino acid identities for full length, extracellular and, cytoplasmic domain comparisons are based on huFcRH3. Closest cytoplasmic relatives are indicated in parentheses. Most identical extracellular comparisons between mouse and human relatives are highlighted in red. Comparisons that are not applicable are left blank.
Figure 8 shows the structural characteristics of the mouse FcRH isoforms.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
The present invention may be understood more readily by reference to the following detailed description of preferred embodiments of the invention and the Examples included therein and to the Figures and their previous and following description.
In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings:
As used in the specification and the appended claims, the singular forms "a,"
"an" and "the" include plural referents unless the context clearly dictates otherwise.
Thus, for example, reference to a receptor includes mixtures of various receptors, reference to "a pharmaceutical carrier" includes mixtures of two or more such carriers, and the like.
Ranges may be expressed herein as from "about" one particular value, and/or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value.
Similarly, when values are expressed as approximations, by use of the antecedent "about,"
it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.
"Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, the phrase "optionally contains two ITAM consensus motifs" means that the two ITAMs may or may not be present and that the description includes both the presence and absence of two ITAM consensus motifs.
As used throughout, by "subject" is meant an individual. Preferably, the subject is a mammal such as a primate, and, more preferably, a human. The term "subject"
can include domesticated animals, such as cats, dogs, etc., livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mouse, rabbit, rat, guinea pig, etc.).
By "isolated nucleic acid" is meant a nucleic acid the structure of which is not identical to that of the naturally occurring nucleic acid or to that of any fragment of the naturally occurring genomic nucleic acid spanning more than three separate genes. The term therefore covers, for example, (a) a DNA which has the sequence of part of the naturally occurring genomic DNA molecules but is not flanked by both of the coding sequences that flank that part of the molecule in the genome of the organism in which it naturally occurs; (b) a nucleic acid incorporated into a vector or into the genomic DNA
of a prokaryote or eukaryote in a manner such that the resulting molecule is not identical to any naturally occurnng vector or genomic DNA; (c) a separate molecule such as cDNA, a genomic fragment, a fragment produced by polymerase chain reaction, or a restriction fragment; and (d) a recombinant nucleotide sequence that is part of a hybrid gene, i.e., a gene encoding a fusion protein.
By "label" is meant any detectable tag that can be attached directly (e.g., a fluorescent molecule integrated into a polypeptide or nucleic acid) or indirectly (e.g., by way of binging to a primary antibody a secondary antibody with an integrated fluorscent molecule) to the molecule of interest. A "label" is any tag that can be visualized with imaging methods. The detectable tag can be a radio-opaque substance, radiolabel, a fluorescent label, or a magnetic label. The detectable tag can be selected from the group consisting of gamma-emitters, beta-emitters, and alpha-emitters, gamma-emitters, positron-emitters, X-ray-emitters and fluorescence-emitters suitable for localization. Suitable fluorescent compounds include fluorescein sodium, fluorescein isothiocyanate, phycoerythrin, and Texas Red sulfonyl chloride. See, de Belder & Wik (Preparation and properties of fluorescein-labelled hyaluronate. Carbohydr.
Res.44(2):251-57 (1975). Those skilled in the art will know, or will be able to ascertain with no more than routine experimentation, other fluorescent compounds that are suitable for labeling the molecule.
Polyneptides The invention provides members of a cluster of FcR and FcR gene relatives encoded by genes in the human chromosome 1q21-23 region, or analogous region in non-human subjects, including for example, chromosome 3 in mouse. A consensus amino acid motif, based on the FcyRI, FcyRII, Fc~yRIII, and pIgR extracellular regions, was used in a GenBank protein database query to identify member of the gene subfamily. Genomic clones were identified that were found to contain FcR
relatives and are termed the Fc receptor homolog (FcRH) subfamily: specifically, FcRHI, FcRH2, FcRH3, and FcRH6. Also, found were mouse Fc receptor homologs designated moFcRl, 2, and 3.
By "homologous" is meant about 25% percent homology or greater. Homology is also characterized by proximity in the location of the genes and by similarities as identified in a composite analysis. As used herein, "percent homology" of two amino acid sequences or of two nucleic acid sequences is determined using the algorithm of Karlin and Altschul (Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990)). Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al.
(J. Mol. Biol. 215:403-410 (1990)). BLAST nucleotide searches are performed with the NBLAST program, score 100, wordlength = 12, to obtain nucleotide sequences homologous to a nucleic acid molecule of the invention. BLAST protein searches are performed with the XBLAST program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to a reference polypeptide . To obtain gapped alignments for comparison purposes, Gapped Blast is utilized as described in Altschul et al. (Nucl.
Acids Res. 25: 3389-3402 (1997)). When utilizing BLAST and Gapped BLAST
programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) are used. See http://ww.ncbi.nlm.nih.gov.
By "FcRH" is meant a Type I transmembrane receptor, or an alternatively spliced form thereof, including, for example, a secreted form or a GPI-anchored form, with homology to the classical Fc receptor family. In a preferred embodiment, the FcRH shows homology with the extracellular regions of Fc~yRI, FcyRII, Fc~yRIII, or pIgR. More specifically, the FcRH shows homology with an amino acid sequence corresponding with the amino terminal sequences of the second Ig domains of the Fc~yRs and the third Ig domain of pIgR or Fc~yRHl. The FcRH can comprise an extracellular region, a transmembrane region, and a cytoplasmic region. The extracellular region preferably comprises one or more Ig domains, and more preferably less than 9, and even more preferably less than 7 or less than 8 Ig domains.
Preferably, the cytoplasmic region comprises more than 107 (including more than 108, 109, 110, 111, 112, 113, 114, 115, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, or 145 amino acids).

Alternatively, the cytoplasmic region comprises less than 104 amino acids (including less than 103, 102, 101, 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80). The cytoplasmic region preferably comprises one or more immunoreceptor tyrosine-based inhibitory or activation motifs ("ITIMs" or "ITAMs).
The invention provides isolated FcRHs (e.g., huFcRH 1, 2, 3, and 6, and moFcRHl-3, as described in detail below), as well as fragments and isoforms thereof.
The isolated amino acid sequences provided herein optionally are combined with a human signal sequence (e.g., MLPRLLLLICAPLCEP (SEQ ID N0:29), MLLWSLLVIFDAVTEQADS (SEQ ID NO:30), MLLWLLLLILTPGREQS (SEQ ID
N0:31), MLLWTAVLLFVPCVG (SEQ ID N0:32)) or a mouse signal sequence (e.g., MPLCLLLLVFAPVGVQS (SEQ ID N0:69), MLPWLLLLICALPCEPA (SEQ ID
N0:72), MSGSFSPCVVFTQMWLTLLVVTPVN (SEQ ID NO:79)).
In one embodiment, the invention provides huFcRHl and its fragments and isoforms. Thus, in one embodiment of the isolated FcRH, the extracellular region comprises less than four Ig domains. Preferably, the cytoplasmic region comprises less than 104 amino acids and, even more preferably, comprises less than 104 and more than 86 amino acids. In one embodiment, the transmembrane region comprises an acidic amino acid (e.g., glutamate or aspartate). The isolated FcRH of the invention comprises a cytoplasmic region having the amino acid sequence of SEQ ID NO:1, in the presence or absence of conservative amino acid substitutions. Further provided is the isolated FcRH, wherein the extracellular region comprises the amino acid sequence of SEQ ID
N0:21, in the presence or absence of conservative amino acid substitutions, and in the presence and absence of a signal sequence. More specifically, the isolated FcRH
comprises the amino acid sequence of SEQ ~ N0:2, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. In one embodiment the signal sequence is MLPRLLLLICAPLCEP (SEQ ID
N0:29). In a preferred embodiment, the FcRH of the invention is expressed by myeloid cells (e.g., granulocytes and monocytes). Additional characteristics of the full length FcRHl include a predicted molecular weight of about 46-47 kDaltons; about 425-(e.g., 429) amino acids in length with about 35 strongly basic(+) amino acids (I~,R), about 45 strongly acidic(-) amino acids (D,E), about 144 hydrophobic amino acids (A,I,L,F,W,V), and about127 polar amino acids (N,C,Q,S,T,I~; a predicted isolelectric point of about 5-5.5 (e.g., 5.310); and charge of about -9 at PH 7Ø

In another embodiment, the invention provides an isolated FcRH corresponding to huFcRH2, its fragments, or isoforms. Thus, the invention provides a FcRH
wherein the cytoplasmic region comprises less than 99 amino acids (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98) and wherein the receptor further 5 comprises an extracellular domain with up to four Ig domains and up to five N-linked glycosylation sites. More specifically, the isolated FcRH has a cytoplasmic region that comprises the amino acid sequence of SEQ ID N0:3, in the presence or absence of conservative amino acid substitutions, or an extracellular region comprising SEQ ID
N0:22, in the presence or absence of conservative amino acid substitutions, and in the 10 presence or absence of a signal sequence. Even more specifically, the isolated FcRH
comprises the amino acid sequence of SEQ ID N0:4, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. In one embodiment, the signal sequence WSLLVIFDAVTEQADS (SEQ ID
N0:30). Additional characteristics of the full length FcRHl include a predicted molecular weight of about 50-60 kDaltons; about 495-515 (e.g., 508) amino acids in length with about 44 strongly basic(+) amino acids (K,R), about 49 strongly acidic(-) amino acids (D,E), about 175 hydrophobic amino acids (A,I,L,F,W,V), and about polar amino acids (N,C,Q,S,T,Y); a predicted isolelectric point of about 6-6.5 (e.g., 6.188); and charge of about -4 at PH 7Ø
In another embodiment, the invention provides huFcRH3, its fragments, and isoforms. More specifically, the invention provides an isolated FcRH having a cytoplasmic region that comprises more than 107 amino acids (e.g., 108, 109, 110, 11 l, 112, 113, 114, 115, 116, 117, 118, 119, 120, 212, 122, 123, 124, 125, 126 ,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 247, 148, 149, 150 amino acids). Optionally, the isolated FcRH has a cytoplasmic region comprising one ITAM and one ITIM. More specifically, the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:S or SEQ ID N0:23, in the presence or absence of conservative amino acid substitutions. In one embodiment, the extracellular domain of the FcRH comprises the amino acid sequence of SEQ ID
NO:24, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. Also provided is an isolated FcRH
comprising the amino acid sequence of SEQ ID NO:6 or SEQ ID N0:25, in the presence or absence of one or more amino acid substitutions, and in the presence or absence of a signal sequence. In one embodiment the signal sequence comprises MLLWLLLLILTPGREQS (SEQ ID N0:31). Additional characteristics of the full length FcRHl include a predicted molecular weight of about 80-90 kDaltons;
about 725-740 (e.g., 734) amino acids in length with about 68 strongly basic(+) amino acids (K,R), about 75 strongly acidic(-) amino acids (D,E), about 232 hydrophobic amino acids (A,I,L,F,W,V), and about 224 polar amino acids (N,C,Q,S,T,Y); a predicted isolelectric point of about 6.5-7.0 (e.g., 6.852); and charge of about -2 at PH 7Ø
The invention further provides an isolated huFcRH6, its fragments, and isoforms. More specifically, the FcRH comprises a cytoplasmic region having the amino acid sequence of SEQ ID N0:26, in the presence or absence or one or more conservative amino acid substitutions. The extracellular domain comprises the amino acid sequence of SEQ ff~ N0:27, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. Also, provided by the invention is a FcRH having the amino acid substitutions of SEQ ID N0:28, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. In one embodiment the signal sequence is MLLWTAVLLFVPCVG (SEQ ff~ N0:32).
The invention further provides a polypeptide comprising the amino acid sequence of SEQ ID N0:1, 21, 2, 3, 22, 4, 5, 23, 24, 6, 25, 26, 27, or 28, in the presence or absence of conservative amino acid substitutions. The invention also provides a polypeptide having at least 80, 85, 90, or 95% homology with SEQ ID NOs: l, 21, 2, 3, 22, 4, 5, 23, 24, 6, 25, 26, 27, or 28.
The invention further provides an isolated moFcRHl isoform, its fragments, and isofornzs. The moFcRHI is an isoform of SEQ 11? N0:68. More specifically, the FcRH comprises four Ig domains, optionally having the sequence of SEQ ff~ N0:
70, in the presence or absence or one or more conservative amino acid substitutions, and in the presence or absence of a signal sequence (e.g., the sequence of SEQ ff~
N0:71).
The invention further provides an isolated moFcRH2, its fragments, and isoforms. The provided isoforms include one isoform with a transmembrane region and one isoform lacking the transmembrane region. More specifically, the FcRH
comprises a cytoplasmic region having the amino acid sequence of SEQ ID N0:76, in the presence or absence or one or more conservative amino acid substitutions. The extracellular domain comprises the amino acid sequence of SEQ ID N0:74, in the presence or absence of conservative amino acid substitutions, and in the presence or absence of a signal sequence. Also, provided by the invention is a FcRH having the amino acid sequence of SEQ ID N0:73, which comprises a transmembrane region, or SEQ ID
N0:77, which lacks the transmembrane region. In each case, the FcRH sequence can include the presence or absence of conservative amino acid substitutions, and the presence or absence of a signal sequence. In one embodiment the signal sequence is the sequence of SEQ ID N0:72.
The invention also provided a moFcRH3, its fragments and isoforms. The cytoplasmic region can comprise the amino acid sequence of SEQ ID N0:81, in the presence or absence of conservative amino acid substitutions. Optionally, the extracellular domain comprises the amino acid sequence of SEQ ff~ N0:80, in the presence or absence of conservative amino acid substitutions or in the presence or absence of a signal sequence (e.g., the sequence of SEQ ID N0:79). The full length sequence optionally has the amin oacid sequence of SEQ m N0:78, in the presence or absence of conservative amino acid substitutions or in the presence or absence of a signal sequence (e.g., the sequence of SEQ ID N0:79).
Fragments, variants, or isoforms of the FcRHs of the invention are provided.
It is understood that these terms include functional variants. Fragments can include the cytoplasmic region, the extracellular region, the transmembrane region or any portion of at least 10 amino acids or any combination of the regions or portions. The variants are produced by making amino acid substitutions, deletions, and insertions, as well as post-translational modifications. Variations in post-translational modifications can include variations in the type or amount of carbohydrate moieties of the protein core or any fragment or derivative thereof. Variations in amino acid sequence may arise naturally as allelic variations (e.g., due to genetic polymorphism) or may be produced by human intervention (e.g., by mutagenesis of cloned DNA sequences), such as induced point, deletion, insertion and substitution mutants. These modifications can result in changes in the amino acid sequence, provide silent mutations, modify a restriction site, or provide other specific mutations.
Amino acid sequence modifications fall into one or more of three classes:
substitutional, insertional or deletional variants. Insertions include amino and/or carboxyl tennii~al fusions as well as intrasequence insertions of single or multiple amino acid residues. Insertions ordinarily will be smaller insertions than those of amino or carboxyl terminal fusions, for example, on the order of one to four residues.
Deletions are characterized by the removal of one or more amino acid residues from the protein sequence. Typically, no more than about 2 to 6 residues are deleted at any one site within the protein molecule. These variants ordinarily are prepared by site-specific mutagenesis of nucleotides in the DNA encoding the protein, thereby producing DNA
encoding the variant, and thereafter expressing the DNA in recombinant cell culture.
Techniques for making substitution mutations at predetermined sites in DNA
having a lcnown sequence are well known and include, for example, M13 primer mutagenesis and PCR mutagenesis. Amino acid substitutions are typically of single residues but may include multiple substitutions at different positions; insertions usually will be on the order of about from 1 to 10 amino acid residues but can be more; and deletions will range about from 1 to 30 residues, but can be more. Deletions or insertions preferably are made in adjacent pairs, i.e. a deletion of 2 residues or insertion of 2 residues.
Substitutions, deletions, insertions or any combination thereof may be combined to arrive at a final construct. The mutations must not place the sequence out of reading frame and preferably will not create complementary regions that could produce secondary mRNA structure. Substitutional variants are those in which at least one residue has been removed and a different residue inserted in its place. Such substitutions generally are made in accordance with Table 1 and are referred to as conservative substitutions.
TABLE l: Amino Acid Substitutions Original ResidueExemplary Substitutions Ala Ser Arg Lys Asn Gln Asp Glu Cys S er Gln Asn Glu Asp Gly Pro His Gln Tle Leu; Val Leu Ile; Val Lys Arg; Gln Met Leu; Ile Phe Met; Leu; Tyr Ser T~' T~. Ser Trp Tyr Tyt. Trp; Phe Val Ile; Leu Substantial changes in function or immunological identity are made by selecting substitutions that are less conservative than those in Table 1, i.e., selecting residues that differ more significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site or (c) the bulk of the side chain. The substitutions that in general are expected to produce the greatest changes in the protein properties will be those in which (a) a hydrophilic residue, e.g. seryl or threonyl, is substituted for (or by) a hydrophobic residue, e.g.
leucyl, isoleucyl, phenylalanyl, valyl or alanyl; (b) a cysteine or proline is substituted for (or by) any other residue; (c) a residue having an electropositive side chain, e.g., lysyl, arginyl, or histidyl, is substituted for (or by) an electronegative residue, e.g., glutamyl or aspartyl; or (d) a residue having a bulky side chain, e.g., phenylalanine, is substituted for (or by) one not having a side chain, e.g., glycine, in this case, (e) by increasing the number of sites for sulfation and/or glycosylation.
Substitutional or deletional mutagenesis can be employed to insert sites for N-glycosylation (Asn-X-Thr/Ser) or O-glycosylation (Ser or Thr). Deletions of cysteine or other labile residues also may be desirable. Deletions or substitutions of potential proteolysis sites, e.g. Arg, is accomplished for example by deleting one of the basic residues or substituting one by glutaminyl or histidyl residues.
Certain post-translational derivatizations are the result of the action of recombinant host cells on the expressed polypeptide. Glutaminyl and asparaginyl residues are frequently post-translationally deamidated to the corresponding glutamyl and asparyl residues. Alternatively, these residues are deamidated under mildly acidic conditions. Other post-translational modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of Beryl or threonyl residues, methylation of the o-amino groups of lysine, arginine, and histidine side chains (T.E.
Creighton, Proteins: Structure and Molecular Properties, W. H. Freeman & Co., San Francisco pp 79-86 [1983]), acetylation of the N-terminal amine and, in some instances, amidation of the C-terminal carboxyl. Modifications in the FcRH can also include modifications in glyco sylation.

In all mutational events, it is understood that the controlling aspect of the mutation is the function that the subsequent protein possesses. The preferred mutations are those that do not detectably change the desired function or that increase the desired function.
Nucleic Acids Also provided is an isolated nucleic acid that encodes the FcRH of the invention. The nucleic acid can be single or double stranded and can be RNA or DNA.
More specifically, the invention provides an isolated nucleic acid, comprising a 10 nucleotide sequence that encodes SEQ ID NO:1, SEQ ID N0:21, SEQ ID N0:2.
SEQ
ID NO: 3, SEQ ID N0:22, SEQ ID N0:4, SEQ ID NO:S, SEQ ID N0:23, SEQ ID
N0:24, SEQ ID N0:25, SEQ ID N0:26, SEQ ID N0:27, SEQ ID N0:28, or SEQ ID
N0:6, SEQ 1T7 N0:70, SEQ ID N0:73, SEQ ID N0:74, SEQ ID N0:76, SEQ ID
N0:77, SEQ ID N0:78, SEQ ID N0:80, SEQ ID N0:81, optionally with conservative 15 amino acid substitutions. Optionally the nucleic acid further encodes a signal sequence (e.g., the signal sequences of SEQ >D N0:29, 30, 31, 32, 71, 75, 79). The isolated nucleic acid optionally encodes the sequences with 80, 85, 90, or 95 %
identity. More specifically, the invention provides an isolated nucleic acid, comprising a nucleotide sequence of SEQ ID N0:7, SEQ ID N0:13, SEQ ID N0:8, SEQ ID N0:34, SEQ ID
N0:9, SEQ ~ N0:14, SEQ ID NO:10, SEQ ID N0:36, SEQ ID NO:11, SEQ ID
NO:15, SEQ ID N0:16, SEQ >D N0:12, SEQ ID N0:38, SEQ ID N0:17, SEQ ID
N0:18, SEQ ID N0:19, SEQ ID N0:20; SEQ m N0:40, SEQ ID N0:84, SEQ ID
N0:85, SEQ 11? N0:87, SEQ m N0:88, SEQ ID N0:89~, SEQ ff~ N0:90, SEQ ID
N0:91, SEQ ID N0:92, SEQ ID N0:93, SEQ ID N0:94, SEQ ID N0:95, SEQ ~
N0:96, SEQ ID N0:97, SEQ ID N0:98, SEQ ID N0:99, SEQ ID NO:100, SEQ ID
NO:101, or SEQ ID N0:102 . Optionally, the isolated nucleic acid can further included bases that encode a signal sequence and thus the nucleotide sequence encoding the extracellular region or full-length huFcRHl, 2, 3, or 6 can optionally further comprise the nucleotide sequence of SEQ ID N0:33, SEQ ID N0:35, SEQ 1177 N0:37, SEQ m N0:39. Optionally, the isolated nucleic acids for rnoFcRHs include nucleic acid sequences that encode signal sequences as well, including for example, those portions of nucleic acid sequences SEQ ID NO:101, SEQ D7 N0:97, SEQ m N0:94, SEQ ID
N0:91, SEQ ID N0:88, SEQ ID N0:84.

Preferably the nucleic acid that encodes the full length FcRHl includes about 1290 bases. The nucleic acid that encodes the full length FcRH2 includes about bases, and the nucleic acid that encodes the full length FcRH3 includes about bases.
The invention also provides an isolated nucleic acid comprising a sequence that hybridizes under stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ff~ N0:7, SEQ m N0:13, SEQ m N0:8, SEQ m N0:34, SEQ m N0:9, SEQ m N0:14, SEQ ff~ NO:10, SEQ m N0:36, SEQ m NO:11, SEQ m NO:15, SEQ m NO:16, SEQ m N0:12, SEQ
m N0:38, SEQ m N0:17, SEQ m N0:18, SEQ m N0:19, and SEQ m N0:20; SEQ
ff~ N0:40, SEQ ID N0:84, SEQ m N0:85, SEQ m N0:87, SEQ m N0:88, SEQ m N0:89, SEQ m NO:90, SEQ m N0:91, SEQ ff~ NO:92, SEQ m N0:93, SEQ m N0:94, SEQ B7 N0:95, SEQ m N0:96, SEQ m N0:97, SEQ m N0:98, SEQ m NO:99, SEQ m NO:100, SEQ m NO:101, or SEQ m N0:102, or the complement of either sequence.
Further provided is a single stranded nucleic acid that hybridizes under stringent conditions to a nucleic acid having the sequence of SEQ m NO:7, SEQ m N0:13, SEQ m N0:8, SEQ m NO:34, SEQ ~ N0:9, SEQ m N0:14, SEQ m NO:10, SEQ
m NO:36, SEQ ff) NO:11, SEQ m NO:15, SEQ m N0:16, SEQ ff~ N0:12, SEQ m N0:38, SEQ ~ N0:17, SEQ m NO:18, SEQ ff~ N0:19, and SEQ m N0:20; SEQ m N0:40, SEQ m N0:84, SEQ m N0:85, SEQ m NO:87, SEQ m N0:88, SEQ m N0:89, SEQ m N0:90, SEQ m N0:91, SEQ ~ N0:92, SEQ m N0:93, SEQ m N0:94, SEQ m N0:95, SEQ m N0:96, SEQ m N0:97, SEQ m N0:98, SEQ m N0:99, SEQ m NO:100, SEQ m NO:101, or SEQ m N0:102.
By "hybridizing under stringent conditions" or "hybridizing under highly stringent conditions" is meant that the hybridizing portion of the hybridizing nucleic acid, typically comprising at least 15 (e.g., 20, 25, 30, or 50 nucleotides), hybridizes to all or a portion of the provided nucleotide sequence under stringent conditions. The term "hybridization" typically means a sequence driven interaction between at least two nucleic acid molecules, such as a primer or a probe and a gene. Sequence driven interaction means an interaction that occurs between two nucleotides or nucleotide analogs or nucleotide derivatives in a nucleotide specific manner. For example, G
interacting with C or A interacting with T are sequence driven interactions.
Typically sequence driven interactions occur on the Watson-Crick face or Hoogsteen face of the nucleotide. The hybridization of two nucleic acids is affected by a number of conditions and parameters known to those of skill in the art. For example, the salt concentrations, pH, and temperature of the reaction all affect whether two nucleic acid molecules will hybridize. Generally, the hybridizing portion of the hybridizing nucleic acid is at least 80%, for example, at least 90%, 95%, or 98%, identical to the sequence of or a portion of a nucleic acid encoding an FcRH of the invention, or its complement.
Hybridizing nucleic acids of the invention can be used, for example, as a cloning probe, a primer (e.g., for PCR), a diagnostic probe, or an antisense probe.
Hybridization of the oligonucleotide probe to a nucleic acid sample typically is performed under stringent conditions. Nucleic acid duplex or hybrid stability is expressed as the melting temperature or Tm, which is the temperature at which a probe dissociates from a target DNA. This melting temperature is used to define the required stringency conditions. If sequences are to be identified that are related and substantially identical to the probe, rather than identical, then it is useful to first establish the lowest temperature at which only homologous hybridization occurs with a particular concentration of salt (e.g., SSC or SSPE). Assuming that a 1% mismatch results in a 1°C decrease in the Tm, the temperature of the final wash in the hybridization reaction is reduced accordingly (for example, if sequence having >95% identity with the probe are sought, the final wash temperature is decreased by 5 °C). In practice, the change in Tm can be between 0.5 °C and 1.5 °C per 1% mismatch. Stringent conditions involve hybridizing at 68 °C in Sx SSC/Sx Denhardt's solution/1.0% SDS, and washing in 0.2x SSC/0.1% SDS at room temperature. Moderately stringent conditions include washing in 3x SSC at 42 °C. The parameters of salt concentration and temperature can be varied to achieve the optimal level of identity between the probe and the target nucleic acid.
Additional guidance regarding such conditions is readily available in the art, for example, in Sambrook et al., 1989, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, NY; and Ausubel et al. (eds.), 1995, Current Protocols in Molecular Biology, (John Wiley & Sons, NY) at Unit 2.10.
The nucleic acids of the present invention are optionally labeled, directly or indirectly. Such labeled nucleic acids are useful in various diagnostic techniques including for example, izz situ hybridization, FISH, izz situ PCR, and PRINS.
Both methods involve the preparation of short sequences of single-stranded nucleic acid probes that are complementary to the nucleic acid sequences that encode an FcRH.
See, e.g., M Andreeff and D Pinkel (1999), An Introduction to Flourescent In-Situ Hybridization: Principles and Clinical Applications, John Wiley ~ Sons, Ltd;
Roche Applied Sciences (2000), Nonradioactive In Situ Hybridization Application Manual;
Roche Applied Sciences (1999), PCR Manual, 2d edition, which are incorporated in their entirety for methods of using nucleic acids.
Vectors cells and methods of using Also provided is an expression vector comprising a nucleic acid of the invention, wherein the nucleic acid is operably linked to an expression control sequence. A wide variety of expression system/regulatory sequence combinations may be employed in expressing the disclosed. Such useful regulatory sequences include, for example, the early or late promoters of SV40, CMV, vaccinia, polyoma or adenovirus, the lac system, the trp system, the TAC system, the TRC system, the LTR
system, the major operator and promoter regions of phage lambda, the control regions of fd coat protein, the promoter for 3-phosphoglycerate kinase or other glycolytic enzymes, the promoters of acid phosphatase (for example, PhoS), the AOX 1 promoter of methylotrophic yeast, the promoters of the yeast a-mating factors, and other sequences known to control the expression of genes of prokaryotic or eukaryotic cells or their viruses, and various combinations thereof.
Such an expression vector can be designed to be expressed by eukaryotic cells or prokaryotic cells. The vectors of the present invention thus provide DNA
molecules which are capable of integration into a prokaryotic or eukaryotic chromosome and expression. The inserted genes in viral and retroviral vectors usually contain promoters, and/or enhancers to help control the expression of the desired gene product. A
promoter is generally a sequence or sequences of DNA that function when in a relatively fixed location in regard to the transcription start site. A
promoter contains core elements required for basic interaction of RNA polymerase and transcription factors, and may contain upstream elements and response elements. It has been shown that all specific regulatory elements can be cloned and used to construct expression vectors that are selectively expressed in specific cell types. For example, the glial fibrillary acetic protein (GFAP) promoter has been used to selectively express genes in cells of glial origin. Expression vectors used in eukaryotic host cells (e.g., yeast, fungi, insect, plant, animal, human or nucleated cells) may also contain sequences necessary for the ternzination of transcription which may affect mRNA expression. These regions are transcribed as polyadenylated segments in the untranslated portion of the mRNA
encoding tissue factor protein. The 3' untranslated regions also include transcription termination sites. It is preferred that the transcription unit also contain a polyadenylation region. One benefit of this region is that it increases the likelihood that the transcribed unit will be processed and transported like mRNA. The identification and use of polyadenylation signals in expression constl-ucts is well established. It is preferred that homologous polyadenylation signals be used in the transgene constructs.
In certain transcription units, the polyadenylation region is derived from the SV40 early polyadenylation signal and consists of about 400 bases. It is also preferred that the transcribed units contain other standard sequences alone or in combination with the above sequences improve expression from, or stability of, the construct.
The invention further provides transfer vectors, which include any nucleotide construction used to deliver genes into cells (e.g., a plasmid), or as part of a general strategy to deliver genes, e.g., as part of recombinant retrovirus or adenovirus (Ram et al. Cancer Res. 53:83-88, (1993)). As used herein, plasmid or viral vectors are agents that transport the disclosed nucleic acids into the cell without degradation and include a promoter yielding expression of the gene in the cells into which it is delivered. In some embodiments the FcRHs are derived from either a virus or a retrovirus. Viral vectors include, for example, Adenovirus, Adeno-associated virus, Herpes virus, Vaccinia virus, Polio virus, AIDS virus, neuronal trophic virus, Sindbis and other RNA
viruses, including these viruses with the HIV backbone. Also preferred are any viral families that share the properties of these viruses that make them suitable for use as vectors.
Retroviruses include Murine Maloney Leukemia virus, MMLV, and retroviruses that express the desirable properties of MMLV as a vector. Retroviral vectors are able to carry a larger genetic payload, i.e., a transgene or marker gene, than other viral vectors, and for this reason are a commonly used vector. However, they are not as useful in non-proliferating cells. Adenovirus vectors are relatively stable and easy to work with, have high titers, and can be delivered in aerosol formulation, and can transfect non-dividing cells. Pox viral vectors are large and have several sites for inserting genes, they are thermostable and can be stored at room temperature. A preferred embodiment is a viral vector which has been engineered so as to suppress the immune response of the host organism, elicited by the viral antigens.
Viral vectors can have higher transaction (ability to introduce genes) abilities than chemical or physical methods to introduce genes into cells. Typically, viral 5 vectors contain, nonstructural early genes, structural late genes, an RNA
polymerase III
transcript, inverted terminal repeats necessary for replication and encapsidation, and promoters to control the transcription and replication of the viral genome.
When engineered as vectors, viruses typically have one or more of the early genes removed and a gene or gene/promotor cassette is inserted into the viral genome in place of the 10 removed viral DNA. Constructs of this type can carry up to about 8 kb of foreign genetic material. The necessary functions of the removed early genes are typically supplied by cell lines that have been engineered to express the gene products of the early genes in trans.
A retrovirus is an animal virus belonging to the virus family of Retroviridae, 15 including any types, subfamilies, genus, or tropisms. Retroviral vectors, in general, are described by Verma, LM., Retroviral vectors for gene transfer. In Microbiology-1985, American Society for Microbiology, pp. 229-232, Washington, (1985), which is incorporated by reference herein. Examples of methods for using retroviral vectors for gene therapy are described in U.S. Patent Nos. 4,868,116 and 4,980,286; PCT
20 applications WO 90/02806 and WO 89/07136; and Mulligan, (Science 260:926-(1993)); the teachings of which are incorporated herein by reference. A
retrovirus is essentially a package which has packed into it nucleic acid cargo. The nucleic acid cargo carnes with it a packaging signal, which ensures that the replicated daughter molecules will be efficiently packaged within the package coat. In addition to the package signal, there are a number of molecules that are needed in cis, for the replication, and packaging of the replicated virus. Typically a retroviral genome, contains the gag, pol, and env genes which are involved in the making of the protein coat. It is the gag, pol, and env genes which are typically replaced by the foreign DNA
that it is to be transferred to the target cell. Retrovirus vectors typically contain a packaging signal for incorporation into the package coat, a sequence which signals the start of the gag transcription unit, elements necessary for reverse transcription, including a primer binding site to bind the tRNA primer of reverse transcription, terminal repeat sequences that guide the switch of RNA strands during DNA
synthesis, a purine rich sequence 5' to the 3' LTR that serve as the priming site for the synthesis of the second strand of DNA synthesis, and specific sequences near the ends of the LTRs that enable the insertion of the DNA state of the retrovirus to insert into the host genome. The removal of the gag, pol, and env genes allows for about 8 kb of foreign sequence to be inserted into the viral genome, become reverse transcribed, and upon replication be packaged into a new retroviral particle. This amount of nucleic acid is sufficient for the delivery of a one to many genes depending on the size of each transcript. It is preferable to include either positive or negative selectable markers along with other genes in the insert.
Since the replication machinery and packaging proteins in most retroviral vectors have been removed (gag, pol, and envy, the vectors are typically generated by placing them into a packaging cell line. A packaging cell line is a cell line that has been transfected or transformed with a retrovirus that contains the replication and packaging machinery, but lacks any packaging signal. When the vector carrying the DNA of choice is transfected into these cell lines, the vector containing the gene of interest is replicated and packaged into new retroviral particles, by the machinery provided in cis by the helper cell. The genomes for the machinery are not packaged because they lack the necessary signals.
The construction of replication-defective adenoviruses has been described (Berkner et al., J. Virology 61:1213-1220 (1987); Massie et al., Mol. Cell.
Biol. 6:2872-2883 (1986); Haj-Ahmad et al., J. Virology 57:267-274 (1986); Davidson et al., J.
Virology 61:1226-1239 (1987); Zhang, Generation and identification of recombinant adenovirus by liposome-mediated transfection and PCR analysis, BioTechniques 15:868-872 (1993)). The benefit of the use of these viruses as vectors is that they are limited in the extent fo which they can spread to other cell types, since they can replicate within an initial infected cell, but are unable to form new infectious viral particles. Recombinant adenoviruses have been shown to achieve high efficiency gene transfer after direct, in vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS parenchyma and a number of other tissue sites (Morsy, J.
Clin.
Invest. 92:1580-1586 (1993); Kirshenbaum, J. Clin. Invest. 92:381-387 (1993);
Roessler, J. Clin. Invest. 92:1085-1092 (1993); Moullier, Nature Genetics 4:154-159 (1993); La Salle, Science 259:988-990 (1993); Gomez-Foix, J. Biol. Chem.
267:25129-25134 (1992); Rich, Human Gene Therapy 4:461-476 (1993); Zabner, Nature Genetics 6:75-83 (1994); Guzman, Circulation Research 73:1201-1207 (1993);
Bout, Human Gene Therapy 5:3-10 (1994); Zabner, Cell 75:207-216 (1993);
Caillaud, Eur. J. Neuroscience 5:1287-1291 (1993); and Ragot, J. Gen. Virology 74:501-(1993)). Recombinant adenoviruses achieve gene transduction by binding to specific cell surface receptors, after which the virus is internalized by receptor-mediated endocytosis, in the same manner as wild type or replication-defective adenovirus (Chardonnet and Dales, Virology 40:462-477 (1970); Brown and Burlingham, J.
Virology 12:386-396 (1973); Svensson and Persson, J. Virology 55:442-449 (1985);
Seth, et al., J. Virol. 51:650-655 (1984); Seth, et al., Mol. Cell. Biol.
4:1528-1533 (1984); Varga et al., J. Virology 65:6061-6070 (1991); Wickham et al., Cell 73:309-319 (1993)).
A viral vector can be one based on an adenovirus which has had the E1 gene removed and these virons are generated in a cell line such as the human 293 cell line.
In another preferred embodiment both the El and E3 genes are removed from the adenovirus genome.
Another type of viral vector is based on an adeno-associated virus (AAV). This defective parvovirus is a preferred vector because it can infect many cell types and is nonpathogenic to humans. AAV type vectors can transport about 4 to 5 kb and wild type AAV is known to stably insert into chromosome 19. Vectors which contain this site specific integration property are preferred. An especially preferred embodiment of this type of vector is the P4.1 C vector produced by Avigen, San Francisco, CA, which can contain the herpes simplex virus thymidine kinase gene, HSV-tk, and/or a marker gene, such as the gene encoding the green fluorescent protein, GFP.
In another type of AAV virus, the AAV contains a pair of inverted terminal repeats (ITRs) which flank at least one cassette containing a promoter which directs cell-specific expression operably linked to a heterologous gene. Heterologous in this context refers to any nucleotide sequence or gene which is not native to the AAV or B19 parvovirus.
Typically the AAV and B19 coding regions have been deleted, resulting in a safe, noncytotoxic vector. The AAV ITRs, or modifications thereof, confer infectivity and site-specific integration, but not cytotoxicity, and the promoter directs cell-specific expression. United States Patent No. 6,261,834 is herein incorproated by reference for material related to the AAV vector.

Molecular genetic experiments with large human herpesviruses have provided a means whereby large heterologous DNA fragments can be cloned, propagated and established in cells permissive for infection with herpesviruses (Sun et al., Nature genetics 8: 33-41, 1994; Cotter and Robertson, Curr Opin Mol Ther 5: 633-644, 1999).
These large DNA viruses (herpes simplex virus (HSV) and Epstein-Barr virus (EBV), have the potential to deliver fragments of human heterologous DNA > 150 kb to specific cells. EBV recombinants can maintain large pieces of DNA in the infected B-cells as episomal DNA. Individual clones carried human genomic inserts up to 330 kb appeared genetically stable The maintenance of these episomes requires a specific EBV
nuclear protein, EBNAl, constitutively expressed during infection with EBV.
Additionally, these vectors can be used for transfection, where large amounts of protein can be generated transiently in vitro. Herpesvirus amplicon systems are also being used to package pieces of DNA > 220 kb and to infect cells that can stably maintain DNA as episomes. Other useful systems include, for example, replicating and host-restricted non-replicating vaccinia virus vectors.
The invention also provides an isolated cell comprising a vector of the invention. The isolated cell can be either a eukaryotic or prokaryotic cell, such as strains of E. coli, Pseudornonas, Bacillus , S'tr~eptomyces; fungi such as yeasts (Saechaf°orrayces , and methylotrophic yeast such as Piclaia, Gandida, Hansenula, and Torulopsis); and animal cells, such as CHO, Rl.l, B-W and LM cells, African Green Monkey kidney cells (for example, COS 1, COS 7, BSC1, BSC40, and BMT10), insect cells (for example, Sue), and human cells and plant cells in tissue culture.
Also provided is a method of making a FcRH, or a fragment or variant thereof comprising culturing a cell comprising a vector of the invention under conditions permitting expression of the FcRH. The method comprises culturing a cell comprising an exogeneous nucleic acid that encodes the FcRH, fragment, or variant, wherein the exogeneous nucleic acid is operably linked to an expression control sequence, and wherein the culture conditions permit expression of the FcRH, fragment, or variant under the control of the expression control sequence; harvesting the medium from the cultured cells, and isolating the FcRH, fragment, or varinat from the cell or culture medium. Optionally the exogenous nucleic acid is the nucleotide sequence of SEQ m N0:7, SEQ ID N0:13, SEQ ID N0:8, SEQ ID N0:34, SEQ ID N0:9, SEQ ID N0:14, SEQ ID NO:10, SEQ ID N0:36, SEQ m NO:11, SEQ ID NO:15, SEQ m N0:16, SEQ

ID N0:12, SEQ ID N0:38, SEQ )D N0:17, SEQ m N0:18, SEQ ID N0:19, and SEQ
)D N0:20; SEQ ID N0:40, SEQ )D N0:84, SEQ ID N0:85, SEQ )D N0:87, SEQ 1D
N0:88, SEQ )D N0:89, SEQ )D N0:90, SEQ m N0:91, SEQ 117 N0:92, SEQ m N0:93, SEQ )D N0:94, SEQ )D N0:95, SEQ m N0:96, SEQ >D N0:97, SEQ )D
N0:98, SEQ ff~ N0:99, SEQ m NO:100, SEQ )D NO:101, or SEQ )D N0:102 or a combination thereof. Optionally, the exogenous nucleic acid further comprises a nucleotide sequence that encodes a signal sequence. In the recombinant methods, the cell can be any known host cell, including for example, a prokaryotic or eukaiyotic cell.
The nucleic acids that are delivered to cells, generally in a plasmid or other vector, typically contain expression controlling systems. For example, the inserted genes in viral and retroviral systems usually contain promoters, and/or enhancers to help control the expression of the desired gene product.
Those skilled in the art of molecular biology will understand that a wide variety of expression systems may be used to produce recombinant FcRH polypeptides (as well as fragments, fusion proteins, and amino acid sequence variants with therapeutic activity) for use in the methods of the invention. Thus, FcRH may be produced using prokaryotic host cells (e.g., Escherichia coli) or eukaryotic host cells (e.g., Saccharomyces cerevisiae, insect cells such as Sf~ cells, or mammalian cells such as CHO cells, COS-1, NIH 3T3, or HeLa cells). These cells are commercially available from, for example, the American Type Culture Collection, Rockville, MD (see also F.
Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, New York, NY, 1998). The method of transformation and the choice of expression vector will depend on the host system selected. Transformation and transfection methods are described, e.g., in Ausubel et al., supra, and expression vectors may be chosen from the numerous examples known in the art.
A nucleic acid sequence encoding an FcRH is introduced into a plasmid or other vector, which is then used to transform living cells. Constructs in which a cDNA
containing the entire FcRH coding sequence, a fragment of the FcRH coding sequence, amino acid variations of the FcRH coding sequence, or fusion proteins of the aforementioned, inserted in the correct orientation into an expression plasmid, may be used for protein expression. In some cases, for example, it may be desirable to express the FcRH coding sequence under the control of an inducible or tissue-specific promoter.

Eukaryotic expression systems permit appropriate post-translational modifications to expressed proteins. Thus, eukaryotic, and more preferably mammalian expression systems, allow glycosylations patterns comparable to naturally expressed FcRH. Transient transfection of a eukaryotic expression plasmid allows the transient 5 production of FcRH by a transfected host cell. FcRH may also be produced by a stably-transfected mammalian cell line. A number of vectors suitable for stable transfection of mammalian cells are available to the public (e.g., see Pouwels et al., Cloning Vectors:
A Laboratory Manual, 1985, Supp. 1987), as are methods for constructing such cell lines (see e.g., F. Ausubel et al., Current Protocols in Molecular Biology, John Wiley &
10 Sons, New York, NY, 1998). Another preferred eukaryotic expression system is the baculovirus system using, for example, the vector pBacPAK9, which is available from Clontech (Palo Alto, CA). If desired, this system may be used in conjunction with other protein expression techniques, for example, the myc tag approach described by Evan et al. (Mol. Cell Biol. 5:3610-3616, 1985) or analogous tagging approaches, e.g., 15 using a hemagluttinin (HA) tag.
Once the recombinant protein is expressed, it can be isolated from the expressing cells by cell lysis followed by protein purification techniques such as affinity chromatography. In this example, an antibody that specifically binds to FcRH, which may be produced by methods that are well-known in the art, can be attached to a 20 column and used to isolate FcRH. Once isolated, the recombinant protein can, if desired, be purified further, e.g., by high performance liquid chromatography (HPLC;
e.g., see Fisher, Laboratory Techniques In Biochemistry And Molecular Biology, Work and Burdon, Eds., Elsevier, 1980).
25 Antibodies The invention also provides a purified antibody or immunologic fragment thereof, wherein the antibody or fragment thereof selectively binds to an FcRH. As used herein, the term "antibody" encompasses, but is not limited to, whole immunoglobulin (i.e., an intact antibody) of any class. Native antibodies are usually heterotetrarneric glycoproteins, composed of two identical light (L) chains and two identical heavy (H) chains. Typically, each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (V(I~) followed by a number of constant domains. Each light chain has a variable domain at one end (V(L)) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains. The light chains of antibodies from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (k) and lambda (1), based on the amino acid sequences of their constant domains.
Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG-1, IgG-2, IgG-3, and IgG-4; IgA-1 and IgA-2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively.
The term "variable" is used herein to describe certain portions of the variable domains that differ in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not usually evenly distributed through the variable domains of antibodies.
It is typically concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a b-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the b-sheet structure. The CDRs in each chain are held together in close proximity by the FR
regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat E. A. et al., "Sequences of Proteins of Ixnmunological Interest" National Institutes of Health, Bethesda, Md. (1987)).
The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

The term "antibody or fragments thereof' can also encompass chimeric antibodies and hybrid antibodies, with dual or multiple antigen or epitope specificities, and fragments, such as F(ab')2, Fab', Fab and the like, including hybrid fragments.
Thus, fragments of the antibodies that retain the ability to bind their specific antigens are provided. For example, fragments of antibodies which maintain FcRH binding activity are included within the meaning of the term "antibody or fragment thereof."
Such antibodies and fragments can be made by techniques known in the art and can be screened for specificity and activity according to the methods set forth in the Examples and in general 'methods for producing antibodies and screening antibodies for specificity and activity (See Harlow and Lane. Antibodies, A Laboratory Manual. Cold Spring Harbor Publications, New York, (1988)).
Also included within the meaning of "antibody or fragments thereof' are conjugates of antibody fragments and antigen binding proteins (single chain antibodies) as described, for example, in U.S. Pat. No. 4,704,692, the contents of which are hereby incorporated by reference.
In one embodiment, the antibody is a monoclonal antibody. The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurnng mutations that may be present in minor amounts. The monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chains) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired activity (See, U.S. Pat. No. 4,816,567 and Morrison et al., Proc.
Natl. Acad. Sci.
USA, 81:6851-6855 (1984)).
Monoclonal antibodies of the invention may be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975) or Harlow and Lane, Antibodies, A Laboratory Manual. Cold Spring Harbor Publications, New York, (1988). In a hybridoma method, a mouse or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent.
Alternatively, the lymphocytes may be immunized in vitro. Preferably, the immunizing agent comprises an FcRH. Traditionally, the generation of monoclonal antibodies has depended on the availability of purified protein or peptides for use as the immunogen.
More recently DNA based immunizations have shown promise as a way to elicit strong immune responses and generate monoclonal antibodies. In this approach, DNA-based immunization can be used, wherein DNA encoding a portion of FcRH, preferably the N- or C- terminal region, is injected into the host animal according to methods known in the art.
Generally, either peripheral blood lymphocytes ("PBLs") are used in methods of producing monoclonal antibodies if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, "Monoclonal Antibodies: Principles and Practice" Academic Press, (1986) pp. 59-103).
Immortalized cell lines are usually transformed mammalian cells, including myeloma cells of rodent, bovine, equine, and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells may be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine ("HAT medium"), which substances prevent the growth of HGPRT-deficient cells.
Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, Calif. and the American Type Culture Collection, Rockville, Md. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (I~ozbor, J.

lmmunol., 133:3001 (1984); Brodeur et al., "Monoclonal Antibody Production Techniques and Applications" Marcel Dekker, Inc., New York, (1987) pp. 51-63).
The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against an FcRH.
Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by innnunoprecipitation or by an ira vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art, and are described further in Harlow and Lane "Antibodies, A Laboratory Manual" Cold Spring Harbor Publications, New York, (1988).
After the desired hybridoma cells are identified, the clones may be subcloned by limiting dilution or FACS sorting procedures and grown by standard methods.
Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells may be grown i~
vivo as ascites in a mammal.
The monoclonal antibodies secreted by the subclones may be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
The monoclonal antibodies may also be made by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, plasmacytoma cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells.
The DNA also may be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567) or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody comprising one antigen-combining site having specificity 5 for FcRH and another antigen-combining site having specificity for a different antigen.
Ira vitro methods are also suitable for preparing monovalent antibodies.
Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art. For instance, digestion can be performed using papain. Examples of papain digestion are described in WO
10 94/29348 published Dec. 22, 1994, U.S. Pat. No. 4,342,566, and Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, (1988). Papain digestion of antibodies typically produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual Fc fragment. Pepsin treatment yields a fragment, called the F(ab')2 fragment, that has 15 two antigen combining sites and is still capable of cross-linking antigen.
The Fab fragments produced in the antibody digestion also contain the constant domains of the light chain and the first constant domain of the heavy chain.
Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain domain including one or more cysteines from the antibody 20 hinge region. The F(ab')2 fragment is a bivalent fragment comprising two Fab' fragments linked by a disulfide bridge at the hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residues) of the constant domains bear a free thiol group. Antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments 25 are also known.
An isolated immunogenically specific epitope or fragment of the antibody is also provided. A specific immunogenic epitope of the antibody can be isolated from the whole antibody by chemical or mechanical disruption of the molecule. The purified fragments thus obtained can be tested to determine their immunogenicity and specificity 30 by the methods taught herein. Irnmunoreactive epitopes of the antibody can also be synthesized directly. An immunoreactive fragment is defined as an amino acid sequence of at least about two to five consecutive amino acids derived from the antibody amino acid sequence.

One method of producing proteins comprising the antibodies of the present invention is to link two or more peptides or polypeptides together by protein chemistry techniques. For example, peptides or polypeptides can be chemically synthesized using currently available laboratory equipment using either Fmoc (9-fluorenylmethyl-oxycarbonyl) or Boc (tef°t -butyloxycarbonoyl) chemistry. (Applied Biosysterns, Inc., Foster City, CA). One skilled in the art can readily appreciate that a peptide or polypeptide corresponding to the antibody of the present invention, for example, can be synthesized by standard chemical reactions. For example, a peptide or polypeptide can be synthesized and not cleaved from its synthesis resin whereas the other fragment of an antibody can be synthesized and subsequently cleaved from the resin, thereby exposing a ternzinal group that is functionally blocked on the other fragment. By peptide condensation reactions, these two fragments can be covalently joined via a peptide bond at their carboxyl and amino termini, respectively, to form an antibody, or fragment thereof. (Grant GA (1992) Synthetic Peptides: A User Guide. W.H. Freeman and Co., N.Y. (1992); Bodansky M and Trost B., Ed. (1993) Principles of Peptide Synthesis.
Springer-Verlag Inc., NY). Alternatively, the peptide or polypeptide can by independently synthesized iri vivo as described above. Once isolated, these independent peptides or polypeptides may be linked to form an antibody or fragment thereof via similar peptide condensation reactions.
For example, enzymatic ligation of cloned or synthetic peptide segments can allow relatively short peptide fragments to be joined to produce larger peptide fragments, polypeptides or whole protein domains (Abrahmsen L et al., Biochemistry, 30:4151 (1991)). Alternatively, native chemical ligation of synthetic peptides can be utilized to synthetically construct large peptides or polypeptides from shorter peptide fragments. This method consists of a two step chemical reaction (Dawson et al.
Synthesis of Proteins by Native Chemical Ligation. Science, 266:776-779 (1994)). The first step is the chemoselective reaction of an unprotected synthetic peptide-oc-thioester with another unprotected peptide segment containing an amino-terminal Cys residue to give a thioester-linked intermediate as the initial covalent product. Without a change in the reaction conditions, this intermediate undergoes spontaneous, rapid intramolecular reaction to form a native peptide bond at the ligation site. Application of this native chemical ligation method to the total synthesis of a protein molecule is illustrated by the preparation ofhuman interleukin 8 (IL-8) (Baggiolini M et al. (1992) FEBS
Lett.

307:97-101; Clark-Lewis I et al., J.Biol.Chem., 269:16075 (1994); Clark-Lewis I et al., Biochemistry, 30:3128 (1991); Rajarathnam K et al., Biochemistry 33:6623-30 (1994)).
Alternatively, unprotected peptide segments can be chemically linked where the bond formed between the peptide segments as a result of the chemical ligation is an unnatural (non-peptide) bond (Schnolzer, M et al. Science, 256:221 (1992)).
This technique has been used to synthesize analogs of protein domains as well as large amounts of relatively pure proteins with full biological activity (deLisle Milton RC et al., Techniques in Protein Chemistry IV. Academic Press, New York, pp. 257-267 (1992)).
The invention also provides fragments of antibodies that have bioactivity. The polypeptide fragments of the present invention can be recombinant proteins obtained by cloning nucleic acids encoding the polypeptide in an expression system capable of producing the polypeptide fragments thereof, such as an adenovirus or baculovirus expression system. For example, one can determine the active domain of an antibody from a specific hybridoma that can cause a biological effect associated with the interaction of the antibody with FcRH. For example, amino acids found to not contribute to either the activity or the binding specificity or affinity of the antibody can be deleted without a loss in the respective activity.
For example, amino or carboxy-terminal amino acids can be sequentially removed from either the native or the modified non-immunoglobulin molecule or the immunoglobulin molecule and the respective activity assayed in one of many available assays. In another example, a fragment of an antibody can comprise a modified antibody wherein at least one amino acid has been substituted for the naturally occurring amino acid at a specific position, and a portion of either amino terminal or carboxy terminal amino acids, or even an internal region of the antibody, has been replaced with a polypeptide fragment or other moiety, such as biotin, which can facilitate in the purification of the modified antibody. For example, a modified antibody can be fused to a maltose binding protein, through either peptide chemistry of cloning the respective nucleic acids encoding the two polypeptide fragments into an expression vector such that the expression of the coding region results in a hybrid polypeptide. The hybrid polypeptide can be affinity purified by passing it over an amylose affinity column, and the modified antibody receptor can then be separated from the maltose binding region by cleaving the hybrid polypeptide with the specific protease factor Xa. (See, for example, New England Biolabs Product Catalog, 1996, pg.
164.).
Similar purification procedures are available for isolating hybrid proteins from eukaryotic cells as well.
The fragments, whether attached to other sequences, can also include insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues, provided the activity of the fragment is not significantly altered or impaired compared to the nonmodified antibody or antibody fragment. These modifications can provide for some additional property, such as to remove or add amino acids capable of disulfide bonding, to increase its bio-longevity, to alter its secretory characteristics, etc. In any case, the fragment must possess a bioactive property, such as binding activity, regulation of binding at the binding domain, etc.
Functional or active regions of the antibody may be identified by mutagenesis of a specific region of the protein, followed by expression and testing of the expressed polypeptide. Such methods are readily apparent to a skilled practitioner in the art and can include site-specific mutagenesis of the nucleic acid encoding the antigen. (Zoller MJ et al. Nucl. Acids Res. 10:6487-500 (1982).
As used herein, the phrase "specific binding" or "selective binding" refers to a binding reaction which is determinative of the presence of the FcRH in a heterogeneous population of proteins and other biologics. Thus, under designated conditions, the antibodies or fragments thereof of the present invention bind to a particular FcRH (e.g., human FcRH 1 or any variant thereof), fragment, or variant thereof and do not bind in a significant amount to other proteins (e.g., human FcRH 2, 3, 4, 5, or 6), present in the subject. 'The absence of binding in the present invention is concisderd to be binding that is less than 1.5 times background (i.e., the level of non-specific binding or slightly above non-specific binding levels), Selective binding to an antibody under such conditions may require an antibody that is selected for its specificity for a particular protein, variant, or fragment. In one embodiment the purified antibody selectively binds to the FcRH comprising a cytoplasmic region with more than 107 or less than 104 amino acids, a transmembrane region, and an extracellular region. More specifically, the antibody in alternative embodiments selectively binds FcRHl but not FcRH2-6; selectively binds FcRH2 but not 1 or 3-6; selectively binds FcRH3 but not FcRHl-2 or 4-6; selectively binds FcRH6 but not 1-5. Thus, as one embodiment, the antibody selectively binds a polypeptide comprising the amino acid sequence of SEQ ID NO:1, 21, or 2, or a subset thereof, but not to polypeptides comprising the amino acid of SEQ ID N0:3, 22, 4, 5, 23, 24, 6, 25, 26, 27, 28, or a subset thereof. In another embodiment the purified antibody binds to the FcRH comprising the amino acid sequence of SEQ ID N0:3, 22, or 4, but not to the FcRH comprising the amino acid of SEQ ID NO:1, 21, 2, 5, 23, 24, 6, 25, 26, 27, or 28.
In yet another embodiment, the purified antibody that binds to the FcRH
comprising the amino acid sequence of SEQ ID NO:S, 23, 24, or 6, but not to the FcRH
comprising the amino acid of SEQ m NO:1, 21, 2, 3, 22, 4, 26, 27, 28. Similarly, the antibodies of the present invention may bind only moFcRHl, but not moFcRH 2 or moFcRH3; may bind only FcRH2 and not FcRHl or FcRH3, and may bind only FcRH3 and not FcRHl or FcRH2.
In certain embodiments, the antibody binds the extracellular region of one or more FcRHs and in other embodiments the antibody binds the cytoplasmic region of one or more FcRHs. In other embodiments the antibody may selectively bind one isoform of a FcRH. For example, the antibody may bind a polypeptide having the amino acid sequence of SEQ 117 N0:23 but not the SEQ ID N0:24 or vice versa.
Furthermore, the antibody can bind to moFcRHl having the amino acid sequence of SEQ LD N0:70, but not to a moFcRHl having amino acid sequence of SEQ ID N0:68.
The antibody may selectively bind a moFcRH2 with a transmembrane region (e.g., having amino acid sequence of SEQ ID N0:73), but not bind to a moFcRH2 lacking a transmembrane region (e.g., having the amino acid sequence of 77). Optionally the antibody of the invention can selectively bind moFcRH but not human, or vice versa.
A variety of immunoassay formats may be used to select antibodies that selectively bind with a particular protein, variant, or fragment. For example, solid-phase ELISA immunoassays are routinely used to select antibodies selectively immunoreactive with a protein, variant, or fragment thereof. See Harlow and Lane.
Antibodies, A Laboratory Manual. Cold Spring Harbor Publications, New York, (1988), for a description of immunoassay formats and conditions that could be used to determine selective binding. The binding affinity of a monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson et al., Anal.
Biochem., 107:220 (1980).
The invention also provides an antibody reagent kit comprising the antibody or fragment thereof of the invention and reagents for detecting binding of the antibody or fragment thereof to a ligand. The kit can further comprise containers containing the antibody or fragment thereof of the invention and containers containing the reagents.
Preferably the ligand is a FcRH, variant, or fragment thereof. Particularly, the kit can detect the presence of one or more FcRHs specifically reactive with the antibody or an 5 immunoreactive fragment thereof. The kit can include an antibody bound to a substrate, a secondary antibody reactive with the antigen and a reagent for detecting a reaction of the secondary antibody with the antigen. Such a kit can be an ELISA kit and can comprise the substrate, primary and secondary antibodies when appropriate, and any other necessary reagents such as detectable moieties, enzyme substrates and color 10 reagents as described above. The diagnostic kit can, alternatively, be an immunoblot kit generally comprising the components and reagents described herein.
Alternatively, the kit could be a radioimmunoassay kit, a Western blot assay kit, an immunohistological assay kit, an immunocytochemical assay kit, a dot blot assay kit, a fluorescence polarization assay kit, a scintillation proximity assay kit, a homogeneous 15 time resolved fluorescence assay kit, or a BIAcore analysis kit.
As used throughout, methods of detecting an FcRH or antigen/antibody complexes, including complexes comprising an FcRH and optionally the antibody of the present invention, can comprise an ELISA (competition or sandwich), a radioimmunoassay, a Western blot assay, an immunohistological assay, an 20 immunocytochemical assay, a dot blot assay, a fluorescence polarization assay (Jolley (1981); Jiskoot et al (1991); Seethala et al. (1998); Bicamumpaka et al.
(1998)), a scintillation proximity assay (Amersham Life Science (1995) Proximity News.
Issue 17;
Amersham Life Science (1995) Proximity News. Issue 18; Park et al. (1999)), a homogeneous time-resolved fluorescence assay (Park et al. (1999); Stenroos et al.
25 (1988); Morrison, 1988)), or a BIAcore analysis Fagerstam et al. (1992) Chromatography 597:397-410. Preferably, the antigei~/antibody complex is detestably tagged either directly or indirectly. Any desired tag can be utilized, such as a fluorescent tag, a radiolabel, a magnetic tag, or an enzymatic reaction product.
Optionally, the antibody or fragment is a humanized antibody or a fully human 30 antibody. For example, the antibodies can also be generated in other species and "humanized" for administration to humans. Alternatively, fully human antibodies can also be made by immunizing a mice or other species capable of making a fully human antibody (e.g., mice genetically modified to produce human antibodies), screening clones that bind FcRH. See, e.g., Lonberg and Huszar (1995) Human antibodies from transgenic mice, Int. Rev. Imrnunol. 13:65-93, which is incorporated herein by reference in its entirety for methods of producing fully human antibodies. As used herein, the term "humanized" and "fully human" in relation to antibodies, relate to any antibody which is expected to elicit a therapeutically tolerable weak immunogenic response in a human subject.
Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab' , F(ab')Z, or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR
of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues.
Humanized antibodies may also comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all or at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR
regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., Nature, 321:522-(1986); Riechmann et al., Nature, 332:323-327 (1988); and Presta, Curr. Op.
Struct.
Biol., 2:593-596 (1992)).
Methods for humanizing non-human antibodies are well known in the art.
Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as "import" residues, which are typically taken from an "import"
variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody.

Accordingly, such "humanized" antibodies are chimeric antibodies (U.S. Pat.
No.
4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important in order to reduce antigenicity.
According to the "best-fit" method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable domain sequences. The human sequence which is closest to that of the rodent is then accepted as the human framework (FR) for the humanized antibody (Sims et al., J.
Immunol., 151:2296 (1993) and Chothia et al., J. Mol. Biol., 196:901 (1987)). Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies (Carte~° et al., Proc. Natl. Acad. Sci.
USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623 (1993)).
It is further important that antibodies be humanized with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, according to a preferred method, humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three dimensional models of the parental and humanized sequences. Three dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the consensus and import sequence so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved. In general, the CDR residues are directly and most substantially involved in influencing antigen binding (see, WO 94/04679 published 3 Mar. 1994).

Transgenic animals (e.g., mice) that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production can be employed. For example, it has been described that the homozygous deletion of the antibody heavy chain joining region (J(I~) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge (see, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551-255 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immuno., 7:33 (1993)). Human antibodies can also be produced in phage display libraries (Hoogenboom et al., J. Mol. Biol., 227:381 (1991); Marks et al., J.
Mol. Biol., 222:581 (1991)). The techniques of Cote et al. and Boerner et al. are also available for the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991)).
In one embodiment, the antibody or fragment thereof is a single chain antibody.
In another embodiment, the antibody or fragment is labeled. Optionally the antibody or fragment is conjugated or fused with a toxin or fragment thereof. Examples of the toxin or toxin moiety include diphtheria, ricin, and modifications thereof.
Dia~,nosis and Treatment The invention provides uses of the reagents described herein in if2 vitro and i~a vivo methods of diagnosing and treating a malignancy of hematopoietic cell lineage or an autoimmune disease in a subject. The reagents of the present invention are also useful in screening for disease manifestations. Such screening may be useful even before the onset of other clinical symptoms and could be used to screening subjects at risk for disease, so that prophylactic treatment can be started before the manifestation of other signs or symptoms.
By "malignancy" is meant a tumor or neoplasm whose cells possess one or more nuclear or cytoplasmic abnormalities, including, for example, high nuclear to cytoplasmic ratio, prominent nucleolar/nucleoli variations, variations in nuclear size, abnormal mitotic figures, or multinucleation. "Malignancies of hematopoietic cell lineage" include, but are not limited to, myelomas, leukemias, lymphomas (Hodgkin's and non-Hodgkin's forms), T-cell malignancies, B-cell malignancies, and lymphosarcomas or other malignancies described in the REAL classification system or the World Health Organization Classification of Hematologic Malignancies. It should be noted that the absence or presence of specific FcRHs can be diagnostic for a particular malignancy of hematopoietic cell linage or can be diagnostic for a particular form of a malignancy (e.g., a specific form of leukemia).
By "inflammatory and autoimmune diseases" illustratively including systemic lupus erythematosus, Hashimoto's disease, rheumatoid arthritis, graft-versus-host disease, Sjogren's syndrome, pernicious anemia, Addison disease, scleroderma, Goodpasture's syndrome, Crohn's disease, autoimmune hemolytic anemia, sterility, myasthenia gravis, multiple sclerosis, Basedow's disease, thrombopenia purpura, insulin-dependent diabetes mellitus, allergy; asthma, atopic disease;
arteriosclerosis;
myocarditis; cardiomyopathy; glomerular nephritis; hypoplastic anemia;
rejection after organ transplantation and numerous malignancies of lung, prostate, liver, ovary, colon, cervix, lymphatic'and breast tissues.
Specifically, the diagnostic methods comprise the steps of contacting a biological sample of the subject with an antibody or nucleic acid of the invention under conditions that allow the antibody to bind to cells of hematopoietic cell lineage or allow the nucleic acid to hybridize, preferably under stringent conditions, with nucleic acids of the biological sample; and detecting the amount or pattern of binding.
Changes in the amount or pattern of binding as compared to binding in a control sample indicate a malignancy or an inflammatory or autoimmune disease.
In various embodiments, the antibody used in the diagnostic method can selectively bind with an FcRH having the amino acid sequence of SEQ ID NO:1, 21, 2, 3, 22, 4, 5, 24, or 6.
The detecting step of the diagnostic method can be selected from methods routine in the art. For example, the detection step can be performed in vivo using a noninvasive medical technique such as radiography, fluoroscopy, sonography, imaging techniques such as magnetic resonance imaging, and the like. In vitro detection methods can be used to detect bound antibody or fragment thereof in an ELISA, RIA, immunohistochemically, FACS, IHC, FISH, or similar assays.
As used throughout, "biological sample" refers to a sample from any organism.
The sample can be, but is not limited to, peripheral blood, plasma, urine, saliva, gastric secretion, feces, bone marrow specimens, primary tumors, embedded tissue sections, frozen tissue sections, cell preparations, cytological preparations, exfoliate samples (e.g., sputum), fine needle aspirations, amnion cells, fresh tissue, dry tissue, and cultured cells or tissue. It is further contemplated that the biological sample of this invention can also be whole cells or cell organelles (e.g., nuclei). The sample can be unfixed or fixed according to standard protocols widely available in the art and can also be embedded in a suitable medium for preparation of the sample. For example, the sample can be embedded in paraffin or other suitable medium (e.g., epoxy or acrylamide) to facilitate preparation of the biological specimen for the detection 10 methods of this invention.
The invention also provides a method of treating a malignancy of hematopoietic cell lineage or an inflammatory or autoimrnune disease in a subject, comprising contacting the subject's malignant cells or inflammatory cells with a therapeutically effective amount of a reagent (e.g., an antibody or nucleic acid) or a therapeutic 15 composition of a reagent of the invention. The contacting step can occur by administration of the reagent or composition using any number of means available in the art. Typically, the reagent or composition is administered to the subject transdermally (e.g., by a transdermal patch or a topically applied cream, ointment, or the like), orally, subcutaneously, intrapulmonaryily, transmucosally, intraperitoneally, 20 intrauterinely, sublingually, intrathecally, intramuscularly, intraarticularly, etc. using conventional methods. In addition, the reagent or composition can be administered via injectable depot routes such as by using 1-, 3-, or 6-month depot injectable or biodegrable materials and methods.
Regardless of the route of administration, the amount of the reagent 25 administered or the schedule for administration will vary among individuals based on age, size, weight, condition to be treated, mode of administration, and the severity of the condition. One skilled in the art will realize that dosages are best optimized by the practicing physician and methods for determining dosage are described, for example in Remington's Pharmaceutical Science, latest edition. Guidance in selecting appropriate 30 doses for antibodies is found in the literature on therapeutic uses of antibodies, e.g., Handbook of Monoclonal Antibodies, Ferrone et al., eds., Noges Publications, Park Ridge, N.J., (1985) ch. 22 and pp. 303-357; Smith et al., Antibodies in Human Diagnosis and Therapy, Haber et al., eds., Raven Press, New York (1977) pp. 365-389. A
typical dose of the antibody used alone might range from about 1 ~g/kg to up to 100 mg/kg of body weight or more per day, and preferably 1 ~g/kg to up to 1 mg/kg, depending on the factors mentioned above. An intravenous injection of the antibody or fragment thereof, for example, could be long-lg of antibody or fragment thereof, and preferably long-lmg depending on the factors mentioned above. For local injection, a typical quantity of antibody ranges from lpg tolmg . Preferably, the local injection would be at an antibody concentration of 1-100 ~g/ml, and preferably 1-20 ~g/ml.
The nucleic acids of the invention can delivered to cells in a variety of ways.
For example, if the nucleic acid of this invention is delivered to the cells of a subject in an adenovirus vector, the dosage for administration of adenovirus to humans can range from about 10~ to 109 plaque forming units (pfu) per injection, but can be as high as 1012 pfu per injection. Ideally, a subject will receive a single injection. If additional injections are necessary, they can be repeated at six month intervals for an indefinite period and/or until the efficacy of the treatment has been established. As set forth herein, the efficacy of treatment can be determined by evaluating the clinical parameters.
The exact amount of the nucleic acid or vector required will vary as described above. Thus, it is not possible to specify an exact amount for every nucleic acid or vector. An appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation given the teachings herein.
The invention further provides a therapeutic composition of the reagent of the invention. Such a composition typically contains from about 0.1 to 90% by weight (such as 1 to 20% or 1 to 10%) of a therapeutic agent of the invention in a pharmaceutically acceptable carrier. Solid formulations of the compositions for oral administration may contain suitable earners or excipients, such as corn starch, gelatin, lactose, acacia, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, calcium carbonate, sodium chloride, or alginic acid. Disintegrators that can be used include, without limitation, microcrystalline cellulose, corn starch, sodium starch, glycolate, and alginic acid. Tablet binders that may be used include acacia, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolindone (PovidoneTM), hydroxypropyl methylcellulose, sucrose, starch, and ethylcellulose. Lubricants that may be used include magnesium stearates, stearic acid, silicone fluid, talc, waxes, oils, and colloidal silica.

Liquid formulations for oral administration prepared in water or other aqueous vehicles may contain various suspending agents such as methylcellulose, alginates, tragacanth, pectin, kelgin, carrageenan, acacia, polyvinylpyrrolidone, and polyvinyl alcohol. The liquid formulations may also include solutions, emulsions, syrups and elixirs containing, together with the active cornpound(s), wetting agents, sweeteners, and coloring and flavoring agents. Various liquid and powder formulations can be prepared by conventional methods for inhalation into the lungs of the mammal to be treated.
Injectable formulations of the compositions may contain various carriers such as vegetable oils, dimethylacetamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, polyols (glycerol, propylene glycol, liquid polyethylene glycol, and the like). For intravenous injections, water soluble version of the compounds may be administered by the drip method, whereby a pharmaceutical formulation containing the antifungal agent and a physiologically acceptable excipient is infused. Physiologically acceptable excipients may include, for example, 5%
dextrose, 0.9% saline, Ringer's solution or other suitable excipients.
Intramuscular preparations, e.g., a sterile formulation of a suitable soluble salt form of the compounds, can be dissolved and administered in a pharmaceutical excipient such as water-for-injection, 0.9% saline, or 5% glucose solution. A suitable insoluble form of the compound may be prepared and administered as a suspension in an aqueous base or a pharmaceutically acceptable oil base, such as an ester of a long chain fatty acid (e.g., ethy; oleate).
A topical semi-solid ointment formulation typically contains a concentration of the active ingredient from about 1 to 20%, e.g., 5 to 10%, in a Garner such as a pharmaceutical cream base. Various formulations for topical use include drops, tinctures, lotions, creams, solutions, and ointments containing the active ingredient and various supports and vehicles. The optimal percentage of the therapeutic agent in each pharmaceutical formulation varies according to the formulation itself and the therapeutic effect desired in the specific pathologies and correlated therapeutic regimens.
The effectiveness of the method of treatment can be assessed by monitoring the patient for known signs or symptoms of the conditions being treated. For example, in the treatment of a malignancy of hematopoietic cell lineage, the reduction or stabilization of the number of abnormally proliferative cells would indicate successful treatment. In the treatment of arthritis, for example, a reduction in the amount of joint inflammation would indicate successful treatment. Thus, by"therapeutically effective"
is meant an amount that provides the desired treatment effect.
The invention further provides a method of modulating a humoral immune response in a subject, comprising administering to the subject an isolated FcRH, an antibody, or nucleic acid of the invention. By "modulation" is meant either up-regulating or down-regulating. Thus, in the case of an allergic response, one skilled in the art would choose to down-regulate the humoral immune response. In the case of exposure of a subject to an infectious agent (e.g., a viral or bacterial agent), one skilled in the art would choose to upregulate the humoral antibody response.
Experimental The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and/or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the invention and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric.

Identification of FcRHl FcRH2, and FcRH3 In order to isolation of FcRH cDNA Clones, rapid amplification of cDNA ends (R.ACE)-PCR was performed by using a Marathon-Ready human lymph node cDNA
library (CLONTECH). Gene-specific primers were as follows: FcRH3, forward 5'-TGAGTCTCAGGGTCACAGTTCCG-3' (SEQ H~ N0:41) and reverse 5'-GCTCTTGAACTTGGATATTTAGGGGT-3' (SEQ ID NO:42); FcRH2, forward 5'-CCAGTGTATGTCAATGTGGGCTCTG-3' (SEQ ID N0:43) and reverse 5'-CGTTGAAAGAGCTCTTGGACTTTTATC-3' (SEQ ID N0:44); and FcRHl, forward 5'-GCCTCAAAAGAAAAATAGGAAGACGTT-3' (SEQ H~ NO:45) and reverse 5'-AAGCTCACATCAGCGACAGGGAC-3' (SEQ ID N0:46). RACE products were subjected to a second round of nested PCR and visualized by agarose gel electrophoresis and ethidium bromide staining.
Primers used in end-to-end amplification to generate full-length cDNAs were as follows: FcRH3, forward 5'-TCTTGGAGATAAGTCGGGCTTT-3' (SEQ ID N0:47) and reverse 5'-ATCCTGCAGCCCAGCCTCGTAGGAG-3' (SEQ ID N0:48); FcRH2, forward 5'-GGTCCTCATGCTGCTGTGGTCATT-3' (SEQ ID N0:49)and reverse 5'-GCTGTTGATCTTCGCTTCTGATTC-3' (SEQ ID NO:50); and FcRHl, forward 5'-ATGCTGCCGAGGCTGTTGCTGTTG3' (SEQ ID NO:51) and reverse 5'-CATAGCATCTTCATAGTCCACATC-3' (SEQ ID N0:52). Each amplification reaction underwent initial denaturation of 94°C for 30 s followed by 30 cycles of denaturation at 94°C for 5 s and annealing at 68°C for 4 min, and final extension at 72°C for 6 min.
PCR products were ligated into the pCR2.1 TOPO T/A vector (Invitrogen).
Inserts were DNA-sequenced on both strands by the dideoxy chain termination method using Thermo Sequenase (Amersham Pharmacia) and an automated sequencer (Li-Cor, Lincoln, NE). Nucleotide and amino acid sequence alignment was analyzed with a DNASTAR (Madison, WI) software package, and homology searches were performed by using BLAST (Altschul, S. F. et al. (1990) J. Mol. Biol. 215, 403-410).
RNA blot analysis was subsequently performed. Northern blots (CLONTECH) were hybridized with 32P-dCTP-labeled probes: a 528-by EcoRI fragment corresponding to the 5' untranslated (UT)-EC1 regions of the FcRH3 cDNA, a 200-by PCR product corresponding to a portion of the 3' UT region of the FcRH2 cDNA, and a 257-by PCR product corresponding to a portion of the 3' UT region of the FcRHl cDNA. Membranes were hybridized for 1 h at 65°C, washed, and exposed to x-ray film (Kubagawa, H. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 5261-5266).
Reverse transcription (RT)-PCR was performed. Human tonsillar cells, obtained with Institutional Review Board approval, were separated into CD19+ and CD19 subpopulations by magnetic cell sorting (Miltenyi Biotec, Auburn, CA). Viable CD19+
cells were stained with FITC-labeled anti-CD38 (Immunotech, Westbrook, ME) and phycoerythrin-labeled anti-IgD mAbs (Southern Biotechnology Associates) before sorting cells with a FACStarPlus instrument (Becton Dickinson) into Trizol reagent (Life Technologies, Grand Island, N~ for RNA isolation. Total cellular RNA was primed with random hexamers and oligo(dT) primers and reverse-transcribed with Superscript II (Life Technologies) into single-stranded cDNA. RT-PCR was performed by using RNA from tonsillar B cells and cell lines, with GIBCOIBRL Taq polymerase (Life Technologies). The following gene-specific primer pairs were used in the RT-5 PCR analysis of FcRHl-5 expresion in cell lines and tonsillar B cell subpopulations:
FcRHl forward, 5'-CTC AAC TTC ACA GTG CCT ACT GGG-3' (SEQ ff~ N0:53) and reverse, 5'-TCC TGC AGA GTC ACT AAC CTT GAG-3' (SEQ ID N0:54);
FcRH2 forward, 5'-CCA GTG TAT GTC AAT GTG GGC TCT G (SEQ ID NO:55) and reverse, 5'-CAT TCT TCC CTC AAA TCT TTA CAC-3' (SEQ ID N0:56); FcRH3 10 forward, 5'-CAG CAC GTG GAT TCG AGT CAC-3' (SEQ ID N0:57) and reverse, 5'-CAG ATC TGG GAA TAA ATC GGG TTG-3' (SEQ ID N0:58) FcRH4 forward, 5'-TCT TCA GAG ATG GCG AGG TCA-3' (SEQ ID N0:59) and reverse, 5'-TTT TGG
GGT GTA CAT CAA CAT ACA AG-3' (SEQ ID N0:60); and FcRH forward, 5'-TGT
TGC CCT GTT TCT TCC AAT ACA-3' (SEQ ID N0:61) and reverse, 5'-CAG AGT
15 TGG CCG ACC TAC GC-3' (SEQ ID N0:62). Each amplification reaction underwent initial denaturation at 94° for 5 min followed by 35 cycles of denaturation at 94° for 30 s, annealing at 60° for 30 s, extension at 72° for 1 min, and final extension at 72° for 7 min. Amplified products were visualized in 1% agarose gels containing ethidium bromide and documented with the Bio-Rad Fluor-S Imager.
20 The following human cell lines were used: REH and Nalm 16 pro-B cell lines (Korsmeyer, S. J. et al. (1983) J. Clin. Invest. 71, 301-313); 697, 207, and OBS pre-B
cell lines (Findley, H. W. et al. (1982) Blood 60, 1305-1309; Martin, D. et al. (1991) J.
Exp. Med. 173, 639-645); Ramos, Daudi, and Raji B cell lines (Pulvertaft, R.
J. V.
(1964) Lancet 1, 238-240; Klein, E. et al. (1968) Cancer Res. 28, 1300-1310;
Klein, G.
25 et al. (1975) Intervirology 5, 319-33431-33); THP-1 and U937 monocytoid cell lines, HL-60 promyelocytic and KG-1 myelocytic cell lines, Jurkat T cell line and the erythroid cell line (American Type Culture Collection).
A consensus sequence was generated that corresponds to the GenBank-derived amino terminal sequences of the second Ig-like domains of 30 FcR (FcyRI and Fc~yRII/III) and the third Ig-like domain of the polymeric Ig receptor:GEPIXLRCHSWK.DKXLXKVTYXQNGKAXI~F'FH (SEQ ID N0:63). A
search of the National Center for Biotechnology Information protein database with this sequence identified two overlapping human genomic bacterial artificial chromosome (BAC) clones, AL135929 and AL356276, which are located at 1q21.2-22. The second clone contained three putative Ig superfamily genes encoding complementary amino acid sequences that were designated FcRHl, FcRH2, and FcRH3. See Figure 1. The predicted amino acid sequences of these gene segments shared 23-57% identity with each other and 14-28% identity with human Fc~yRI (CD64). Further analysis of the FcRH locus led to the identification of two additional genes (FcRH4, and FcRHS) and one pseudogene (FcRH4yr), immediately centromeric of FcRHl-3, two of which have recently been described as IRTAl (FcRH4) and IRTA2 (FcRHS) (Hatzivassiliou, G.
et al. (2001) Immunity 14, 277-289).
To determine whether these genes are expressed by lymphocytes, the predicted amino acid sequences of their protein products were used to search the Lymphochip expressed sequence tag database with the TBLASTN algorithm (Alizadeh, A. A. et al.
(2000) Nature (London) 403, 503-511). Two expressed sequence tags (AA505046 and AA282433) were identified that share complete identity over 23 amino acids in their translated ORFs with the N terminus of FcRHI . Lymphochip microarray data analysis indicated that these expressed sequence tags are expressed at relatively high levels in peripheral lymphoid tissues, including the lymph nodes, tonsils, resting peripheral B
cells, and normal germinal center (GC) B cells. Among the different lymphoid malignancies, their expression proved to be highest in chronic lymphocytic leulcemias, follicular lymphomas, and some diffuse large cell lymphomas of B lineage.
FcRHl, FcRH2, and FcRH3 cDNAs were isolated by RACE-PCR from a human lymph node cDNA library in both 5' and 3' directions. Full-length cDNAs of the coding regions for FcRHl, FcRH2, and FcRH3 were obtained by end-to-end PCR
using unique primers generated from the cDNA sequences delineated for the 5' UT and 3'UT
regions. Southern blot analysis of human genomic DNA digested with Bar~zHI, EcoRI, or Hin.dIII using cDNA probes specific for the 3' UT regions of each cDNA revealed either one or two hybridizing fragments, suggesting that FcRHl, FcRH2, and FcRH3 are encoded by single genes. Analysis of full-length cDNA
sequences indicated that FcRHl, FcRH2, and FcRH3 have ORFs of 1,287 bp, 1,524 bp, and 2,202 bp, respectively, and encode type I transmembrane proteins of 429 aa, 508 aa, and 734 aa, respectively. Based on predicted consensus signal peptide cleavage sites (Von Heijne, G. (1986) Nucleic Acid Res. 14, 4683-4690; Nielsen, H. (1997) Protein Eng. 10, 1-6), the relative core peptide molecular masses were estimated as 45,158 for FcRHl, 53,407 for FcRH2, and 78,849 for FcRH3. These type I transmembrane proteins possess 3-6 extracellular C2 (Williams, A. F. & Barclay, A. N. (1988) Annu.
Rev. Immunol. 6, 381-405; Bork, P. et al. (1994) J. Mol. Biol. 242, 309-320;
Vaughn, D. E. & Bjorkman, P. J. (1996) Neuron 16, 261-273) type Ig-like domains with 3-potential N-linked glycosylation sites, uncharged transmembrane segments, and relatively long cytoplasmic tails containing consensus motifs for ITIMs and/or ITAMs.
See Fig. 2A.
Multiple alignment analysis of the translated cDNAs, using FcRH3 as the index sequence of comparison, indicates that FcRHl, FcRH2, and FcRH3 have highly conserved hydrophobic signal peptides and corresponding Ig-like extracellular domains (Fig. 2B). Their hydrophobic transmembrane (uncharged with the exception of FcRHl which includes an acidic domain) domains (Sonnhammer, E. L. L. et al. (1998) in A
Hidden Markov Model for Predicting Transmembrane Helices in Protein Sequences, eds. Glasgow, J., Littlejohn, T., Major, F., Lathrop, R., Sankoff, D. &
Sensen, C. (Am.
Assoc. for Artificial Intelligence, Menlo Park, CA), pp. 175-182) are also well conserved, but their cytoplasmic domains are not. FcRHl has a long cytoplasmic tail containing three potential ITAMs, the first and third of which fit the consensus sequence (E/D)-X-X-Y-X-X-(L/I)-X~_8-Y-X-X-(L/I) (SEQ ID N0:64, with six amino acid between the consensus sequences; SEQ ID N0:65, with seven amino acid residues between the consensus sequences; and SEQ ID NO:66, with eight amino acid residues between the consensus sequences), whereas, the second has only one tyrosine residue.
The shorter cytoplasmic domain of FcRH2 contains one potential ITAM and two ITIM
consensus sequences (I/V/L/S)-X-Y-X-X-(L/V) (SEQ ID N0:67) separated by 22 amino acids. FcRH3 has the longest cytoplasmic tail. It contains one potential ITAM, one ITIM, and another potential ITAM that also has a single tyrosine residue.
An RNA blot analysis with gene-specific probes was performed on 16 human tissues, including six primary or secondary lymphoid tissues. RNA blots were analyzed with discriminating a32P-dCTP-labeled probes generated from the respective FcRH
cDNAs. The following probes were used: (Top) a PCR-generated, 257-by probe specific to the 3' UT region of FcRHl; (Middle) a PCR-generated, 290-by probe corresponding to the 3' UT region of FcRH2; and (Bottom) a 528-by EcoRI-digested fragment of the 5' end of the FcRH3 cDNA corresponding to its 5' UT region, S
l, S2, and EC 1 domains. The relative mRNA abundance was indicated by (3-actin probe.
All three FcRH gene probes hybridized with transcripts in the secondary lymphoid organs, spleen and lymph node. An FcRHl-specific probe hybridized with spleen and lymph node transcripts of about 3.5 kb and about 6.0 kb. Additional hybridization bands of about 0.7 kb and about 1.5 kb were observed for heart, skeletal muscle, kidney, liver, and, in less abundance, placental tissue. Larger transcripts also were seen in skeletal muscle (about 6.0 kb) and in kidney and placenta (about 4.4 kb). An FcRH2-specific probe hybridized to about 3.0-kb, about 4.4-kb, and about 5.5-kb transcripts most abundantly in spleen and lymph node. A transcript of approximately 2.4-kb was notable in the kidney. An FcRH3 probe hybridized with about 3.5-kb, about 5.5-kb, and about 7.0-kb transcripts chiefly in spleen and lymph node. These also were seen, albeit in lesser abundance, in peripheral blood lymphocytes, thymus, and bone marrow samples.
Additionally, a unique transcript of about 1.35 kb was evident in skeletal muscle. These results indicated expression of FcRHl, FcRH2, and FcRH3 in peripheral lymphoid organs, whereas tissue specific differences in alternative splicing or polyadenylation were suggested by the differential expression of transcripts with variable size in nonlymphoid tissues. RTPCR analysis to date of non-lymphoid tissue skeletal muscle, however, does not reveal transcripts despite the Northern analysis results.
When FcRH expression was examined by RT-PCR analysis of cell lines representing different hematopoietic lineages, FcRHI, FcRH2, and FcRH3 expression was found in every mature B cell line tested (Table 2). FcRH2 and FcRH3 expression was limited to the mature B cell lines and not seen in the other types of cells examined.
In contrast, FcRHl expression was seen in pro-B, T, and myeloid cell lines, although not in an erythroid cell line.
Table 2. Expression of FcRH transcripts in human B cell lines Cell Type Cell lisae FcRIIl FcRH2 FcRH3 Pro-B ~ REH + - -Nahn 16 + -Pre-B ~ 697 - - -OBS - -B ~ Ramos + + +

Daudi + + +

Raji + + +

T Jurkat + - -Monocytic THP-1 + - -Myelomonocytic LT937 + -Promyelocytic HL-60 + - -Myelocytic KG-1 + - -Erythroid K562 - -FcRHl, FcRH2, and FcRH3 expression in cell lines was determined by RT-PCR.
RT-PCR analysis of sorted populations of peripheral blood cells indicated that FcRHl, FcRH2, FcRH3, and FcRHS are expressed at relatively high levels in CD19+ B
cells, whereas FcRH4 was expressed at only trace levels. FcRH3 expression was observed in CD3+ T cells whereas transcripts of FcRHl were barely detectable.
FcRHl expression also was observed in circulating granulocytes.
To refine the analysis of FcRH expression in secondary lymphoid tissues, tonsillar lymphocyte subpopulations were isolated. The five discrete subpopulations of B lineage cells, which can be distinguished by their differential expression of cell surface IgD and CD38, represent different stages in B cell differentiation:
follicular mantle (IgD+CD38), pre-GC (IgD+CD38+), GC (IgDCD38+), memory (IgDCD38), and mature plasma cells (CD382+) (Pascual, V. (1994) J. Exp. Med. 180: 329-339). RT-PCR analysis of FcRHl-5 expression in tonsillar B cell subpopulations was performed.
Viable cells were magnetically sorted into CD19- non-B cells and CD19+ B
cells. The latter were stained with anti-IgD and anti-CD38 mAbs, and the five subpopulations indicated (CD38- IgD-, CD38- IgD+, CD38+ IgD+, CD38+ IgD-, and CD382+) were sorted by flow cytometry. RT-PCR analysis of FcRH transcripts in non-B cells and the B cell subpopulations was also performed. After cDNA preparation, PCR
amplification was performed on the equivalent template of approximately 10 k cells.
Glyceraldehyde-3-phosphate dehydrogenase (GADPH) was amplified as a positive control.
RT-PCR analysis indicated little or no expression of FcRH transcripts in the non-B lineage CD19- cells, most of which are T cells. However, CD19+
subpopulations displayed coordinate expression of FcRHl, FcRH2, and FcRH3 transcripts in follicular mantle, naive, GC, and memory B cell subpopulations, but yielded no evidence of FcRH transcripts in pre-GC B cells or plasma cells. In contrast, FcRH4 transcripts were restricted to the follicular mantle and memory B cells, whereas FcRHS
expression extended to mature plasma cells.
The relationship between the five FcRHs was examined by comparing their full-length, extracellular, and individual Ig-like domain amino acid sequences.
This analysis, which included a recently identified mouse FcRH ortholog (moFeRH) and members of the FcR family, used the CLUSTAL method algorithm (Higgins, D. G. &
Sharp, P. M. (1989) Comput. Appl. Biosci. 5, 151-153). Comparison of the full-length 10 sequences of other FcRH family members with FcRH3 indicated 40-47%
identity. By comparison, the degree of FcRH3 homology with the moFcRH was found to be 35%
and 21-24% with FcR members residing on chromosome 1, FcyRI, FcyRII, Fc~yRIII, and FcsRI. A lower level of amino acid identity (14%) was observed for the chromosome 19 LRC member, FcaR. A slightly higher degree of extracellular homology was 15 evident. Pairwise analysis of the individual Ig-like subunits indicated conservation in membrane-distal to membrane-proximal ordering of extracellular domain composition among family members. Although similar Ig domain subunits were shared among family members, the individual receptors were found to be composed of unique domain combinations. The extracellular domain configuration of the moFcRH most closely 20 resembled that of FcRH2, with which it has 46% identity. The extended pairwise comparison of the FcRH family with known FcRs suggested the conservation of these Ig-like domains to some degree throughout the greater family. The resemblance is particularly evident in the FcRH3 membrane-distal domains that correspond to the three FcyRI domains and the two domains of FcYRII, FcyRIII, and the FcR y-chain.
This 25 analysis suggests the ancestral occurrence of differential duplication and diversification of the individual Ig-like subunits in the respective FcRH family members. The data also indicate that the FcRHs are more similar to their FcR neighbors on chromosome 1 than to their FcR relative on chromosome 19.
The genomic sequence analysis of relevant chromosome 1 q21 BAC
30 clones indicated that the entire FcRH locus spans 300 kb. The FcRH genes lie in the same transcriptional orientation toward the centomere. Exon-intron boundaries were characterized by sequence comparison of their respective cDNA clones and the AG/GT
rule. The FcRHl gene consists of 11 exons and 10 introns spanning about 28 kb.
The first exon, 5' UT/S 1, encodes the 5' UT region, the ATG translation initiation colon, and the first half of a split signal peptide. S2, the second exon, is separated from 5' UT/S1 by a long intron of 12.9 kb and, like the neighboring FcRs, is 21 by in length (van de Winkel, J. G. & Capel, P. J. (1993) Immunol. Today 14, 221; Kulczycki, A., Jr. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 2856-2860; Pang, J.
et al. (1994) J. Immunol. 151, 6166-6174.). The extracellular region is encoded by three closely clustered exons, ECl-EC3, that code for the three Ig-like domains. The membrane-proximal, transmembrane, and the proximal portion of the cytoplasmic domain are encoded by a single sixth exon, TM. The cytoplasmic tail is encoded by five exons, CY1-CYS, and the CYS also encodes the beginning of the 3' UT region.
FcRH2 contains 12 exons and 11 introns that span 30 kb. It also contains two exons that encode a split signal peptide, the first of which, 5'UT/S1, includes the 5' UT
region, the ATG translation initiation colon, and first half of the signal peptide. The second exon, S2, is 21 by in length. Exons 3-6 encode the four extracellular domains, EC1-EC4. The seventh exon encodes the membrane-proximal, transmembrane, and the proximal portion of the cytoplasmic domain. The FcRH2 cytoplasmic tail is encoded by five exons, CY1-CYS, the last exon of which includes the termination of the ORF and beginning of the 3' UT region.
The FcRH3 gene consists of 16 exons and 15 introns that span about 24 kb.
Unlike FcRHl and FcRH2, its 5' UT region is encoded by two exons, 5' UTl and a second, 5'UT2/S 1, that also encodes the ATG translation initiation colon and the beginning of the split signal peptide. The third exon, S2, is also 21 by in length.
Extracellular domains encoded by six exons, EC1-EC6, are followed by exon 10 that encodes the membrane-proximal, transmembrane, and the proximal portion of the cytoplasmic domain. The cytoplasmic tail is encoded by five exons, CY1-CYS;
the last contains the beginning of the 3' UT region.

Identification of HuFcRH6 FcRH6 is located in the midst of the classical FcRs at 1q21-23. Its genomic structure indicates, like the classical FcRs and FcRHI-5, a split hydrophobic signal peptide encoded by two exons the second of which is 2lbp.
FcRH6 was characterized using the methods described in Example 1. A
composite analysis of Ig-like domains for relatedness with the other huFcRHs was performed. See Figure xxx. Sequence analysis of huFcRH6 indicates its type I
transmembrane form contains a consensus motif for a single ITAM, or a single or two ITIM's.
Initial RT-PCR analysis of huFcRH6 in human tissues and cell lines (as described in Example 1) reveals transcript expression in normal tonsil and lymph nodes. In cell lines, expression of huFcRH6 was identified in myeloid cell lines THP-1 (monocytic), U937 (myelomonocytic), and KG-1 (myelocytic). Limited expression if any was identified in the 207 pre-B cell line and the Daudi B cell line.

Generation of Transfectants and Antibodies Recombinant constructs for transfection and stable expression of huFcRHl-5 have been generated. The constructs have been ligated into a CMV driven mammalian expression vector with and without green fluorescent protein (GFP) fusion at the carboxyl terminus. Surface expression of huFcRHI and huFcRH3 was detected for both GFP and non-GFP forms by staining with antibody supernatant. The antibody supernatant was derived from hybridomas generated by mice immunized with recombinant extracellular protein of the respective FcRH. The constructs for huFcRH2, 4, and 5 have been detected by green fluorescence as well as surface expression for FcRH4.
Monoclonal antibodies have been generated, including, for example, an antibody that binds FcRHl. The preliminary analysis of FACS staining for FcRHl expression with monoclonal antibody 1-SA3 labeled with a FITC conjugate (mouse anti human FcRHl) in peripheral blood from normal volunteers indicates virtually all CD19+ B cells have huFcRHl expression, as do CD14+ monocytes and CD13+
granulocytes. CD3+ T cells have limited to no expression of FcRHl. Staining of B-CLL
samples from two different patient peripheral blood samples indicates that virtually all CDS+/CD19+ B-CLL cells are positive for the FcRHl 1-SA3 antigen. By western blot analysis of recombinant protein for FcRHl-5 extracellular regions 1-SA3 appears specific for FcRHl. 1-SA3 also stains B cell lines Daudi and Raji.

Identification of MoFcRHl-3 A family of three mouse Fc Receptor Homologs (MoFCRHs) were identified and cloned. Amino acid sequences from the membrane proximal Ig-like domains of huFcRHl-5 were used to identify putative mouse FcRH orthologs in the NCBI or Cetera genomic, EST, and protein databases using the protein BLAST (BLASTP) and the translated nucleotide BLAST (TBLASTN] algorithms, respectively. The location of moFcR family is split between chromosomes 1 and 3 in regions syntenic with human chromosome 1q21-23. See Figure 4. The mo FcRH are located on mouse Ch3. Approximate positions were determined from Genbank, Cetera, and Mouse Genome Informatics databasesContigs of ESTs were generated to determine the putative cDNA sequences.
Genomic organization was determined by comparing cDNA clones generated from RACE PCR with GenBank and Cetera genomic sequences. DNAStar software was used for analysis of exon-intron boundaries which were characterized by sequence comparison and the AG/GT rule. All three genes contain a split signal sequence with a 2lbp S2 exon (exon 2) which is found in all FcR and huFcRH genes on human chromosone 1.
A comparison of tyrosine based motifs in FcRH cytoplasmic tails indicated homology with the huFcRH family. See Figure 5. An analysis of sequence homology conservation is further shown in Figures 6 and 7.
Expression of the moFcRHs in tissue and cell lines was also characterized as described in Example 1. Briefly, RT-PCR was performed on mouse tissues and cell lines with gene specific primers. Viable tissue was placed in TRIzoI reagent for RNA
extraction. After cDNA preparation PCT amplification was performed on equivalent template amounts. Actin was amplified as a positive control. McFcRH3 appears to have preferential expression in cells of B lineage. The results are shown in Tables 3-4.

Table 3: Tissue Distribution of moFcRH Expression TISSUE MoFcRHl MoFcRH2 MoFcRH3 Bone Marrow + + +

Thymus + + +

Spleen + + +

Lymph Node + + +

Peyer's Patches+ + +

Peripheral + + +
Blood Brain + -Liver + + -Heart + -Muscle + - -Kidney + - -Lung + + -Intestine + + +

Testes I + I -Table 4. Expression of moFcRH transcripts in cell lines Cell Type Cell lirasFcRHI FcRH2 FcRH3 Pro-B SCID7 + +/- +

Raw8.1 + + -Pre-B 70Z/3 + + +

BC76 - ' + +

18-81 + + +

Imm. B WEHI-231 + + +

WEHI-279 + + +

B A20 + + +

X16C8.5 + + +

T EL4 + +/- -/+

NKT NKT + +/- -~T 2C12 + +/- -Myeloid WEHI-3 + - -Lymphoid YAC-1 + + -Fibroblast 3T3 + +/- -Expression in cell lines was determined by RT-PCR.
5 The mouse FcReceptor Homologs include secreted or type I transmembrane isolfrms that have unique cytoplasmic tails with potential activation and inhibition motifs. Their chromosomal location, Ig domain homology, and genomic organization indicate the mouse FcReceptor Homologs are orthologs of the huFcRH that have evolved a significant level of diversity. moFcRHl, moFcRH2, and moFcRH3 are 10 predicted to encode secreted or type I transmembrane proteins based on their amino acid sequences. moFcRHl has two secreted isoforms both of which have extracellular (EC) regions of four Ig-like domains with five potential sites for N-linked glycosylation. One isoform is a fusion protein with a type B scavenger receptor domain containing 8 cysteines. moFcRH2 has secreted and type I isoforms containing two Ig-15 like domains with five N-linked glycosylation sites. The type I isoform has an uncharged transmembrane region which the secreted isoform lacks. Both isoforms contain the cytoplasmic portion which is long in the transmembrane form and contains five tyrosines including a consensus sequence for one potential immunoreceptor tyrosine-based activating motif . moFcRH3 contains five Ig-like domains with six potential sites of N-linked glycosylation. Its transmembrane domain is also uncharged and the cytoplasmic region contains one potential ITAM and one potential immunoreceptor tyrosine-based inhibitory motif. The amino acid (aa) length of individual regions and full length (FL) isoforms, as well as approximate molecular weight (MW) in Daltons (Da), is indicated in the structural diagram of Figure ~.
Throughout this application, various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.
It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.

SEQUENCE LISTING
<110> The UAB Research Foundation Davis, Randall S.
Cooper, Max D.
<120> MEMBERS OF THE FC RECEPTOR HOMOLOG GENE FAMILY (FCRH1-3, 6), RELATED REAGENTS, AND USES THEREOF
<130> 21085.0037P1 <141> 2003-03-25 <150> US 60/367,667 <151> 2002-03-25 <160> 102 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 99 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 1 Lys Arg Lys I1e Gly Arg Arg 5er Ala Arg Asp Pro Leu Arg Ser Leu 1 5 l0 15 Pro Ser Pro Leu Pro Gln Glu Phe Thr Tyr Leu Asn Ser Pro Thr Pro Gly Gln Leu Gln Pro Ile Tyr Glu Asn Val Asn Val Val Ser Gly Asp Glu Val Tyr Ser Leu,Ala Tyr Tyr Asn Gln Pro Glu G1n Glu Ser Val Ala Ala Glu Thr Leu Gly Thr His Met Glu Asp Lys Val Ser Leu Asp Ile Tyr Ser Arg Leu Arg Lys Ala Asn Ile Thr Asp Val Asp Tyr Glu Asp Ala Met <210> 2 <211> 413 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 2 Ala Glu Leu Phe Leu Tle Ala Ser Pro Ser His Pro Thr Glu Gly Ser Pro Val Thr Leu Thr Cys Lys Met Pro Phe Leu Gln Ser 5er Asp Ala Gln Phe Gln Phe Cys Phe Phe Arg Asp Thr Arg Ala Leu Gly Pro Gly Trp Ser Ser Ser Pro Lys Leu Gln Ile Ala Ala Met Trp Lys Glu Asp Thr Gly Ser Tyr Trp Cys Glu Ala Gln Thr Met Ala Ser Lys Val Leu Arg Ser Arg Arg Ser Gln Ile Asn Val His Arg Val Pro Val Ala Asp Val Ser Leu Glu Thr Gln Pro Pro Gly Gly Gln Val Met Glu Gly Asp Arg Leu Val Leu I1e Cys Ser Val Ala Met Gly Thr Gly Asp Ile Thr Phe Leu Trp Tyr Lys Gly Ala Val Gly Leu Asn Leu Gln Ser Lys Thr Gln Arg 5er Leu Thr Ala Glu Tyr Glu Ile Pro Ser Val Arg Glu Ser Asp Ala Glu Gln Tyr Tyr Cys Val Ala Glu Asn Gly Tyr Gly Pro Ser Pro Ser Gly Leu Val Ser Ile Thr Val Arg Ile Pro Val Ser Arg Pro Ile Leu Met Leu Arg Ala Pro Arg Ala Gln Ala Ala Val Glu Asp Val Leu Glu Leu His Cys Glu Ala Leu Arg Gly Ser Pro Pro Ile Leu Tyr Trp Phe Tyr His Glu Asp Ile Thr Leu Gly Ser Arg Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Glu Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asn Asn Gly Leu Gly Ala Gln Arg Ser Glu Ala Val Thr Leu Asn Phe Thr Val Pro Thr Gly Ala Arg Ser Asn His Leu Thr Ser Gly Val Ile Glu Gly Leu Leu Ser Thr Leu Gly Pro Ala Thr Val Ala Leu Leu Phe Cys Tyr Gly Leu Lys Arg Lys Ile Gly Arg Arg Ser Ala Arg Asp Pro Leu Arg Ser Leu Pro Ser Pro Leu Pro Gln Glu Phe Thr Tyr Leu Asn Ser Pro Thr Pro Gly Gln Leu Gln Pro Ile Tyr Glu Asn Val Asn Val Val Ser Gly Asp Glu Val Tyr Ser Leu Ala Tyr Tyr Asn G1n Pro Glu Gln Glu Ser Val Ala Ala Glu Thr Leu Gly Thr His Met ~Glu Asp Lys Val Ser Leu Asp Ile Tyr Ser Arg Leu Arg Lys Ala Asn Ile Thr Asp Val Asp Tyr Glu Asp Ala Met <210> 3 <211> 86 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 3 His Lys Ile Ser Gly Glu Ser Ser A1a Thr Asn Glu Pro Arg Gly Ala Ser Arg Pro Asn Pro Gln Glu Phe Thr Tyr Ser 5er Pro Thr Pro Asp Met Glu Glu Leu Gln Pro Val Tyr Val Asn Val G1y Ser Val Asp Val Asp Val Val Tyr Ser Gln Val Trp Ser Met Gln Gln Pro Glu Ser Ser Ala Asn Ile Arg Thr Leu Leu Glu Asn Lys Asp Ser Gln Val Tle Tyr Ser Ser Val Lys Lys Ser <210> 4 <21l> 489 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 4 Leu Thr Leu Val Ala Pro Ser Ser Val Phe Glu Gly Asp Ser Ile Val Leu Lys Cys Gln Gly Glu Gln Asn Trp Lys Tle Gln Lys Met Ala Tyr His Lys Asp Asn Lys Glu Leu Ser Val Phe Lys Lys Phe Ser Asp Phe Leu Tle Gln Ser Ala Val Leu Ser Asp Ser Gly Asn Tyr Phe Cys Ser Thr Lys Gly Gln Leu Phe Leu Trp Asp Lys Thr Ser Asn Ile Val Lys Ile Lys Val Gln Glu Leu Phe Gln Arg Pro Val Leu Thr Ala Ser Ser Phe Gln Pro Ile Glu Gly Gly Pro Val 5er Leu Lys Cys Glu Thr Arg Leu Ser Pro Gln Arg Leu Asp Val Gln Leu Gln Phe Cys Phe Phe Arg Glu Asn Gln Val Leu Gly Ser Gly Trp Ser Ser Ser Pro Glu Leu Gln Ile Ser Ala Val Trp Ser Glu Asp Thr G1y Ser Tyr Trp Cys Lys Ala Glu Thr Va1 Thr His Arg Ile Arg Lys Gln 5er Leu Gln Ser Gln Ile _ 165 170 175 His Val Gln Arg Ile Pro Ile Ser Asn Val Ser Leu Glu Ile Arg Ala Pro Gly Gly Gln Val Thr Glu Gly Gln Lys Leu Ile Leu Leu Cys Ser Val Ala G1y Gly Thr Gly Asn Val Thr Phe Ser Trp Tyr Arg Glu Ala Thr Gly Thr Ser Met G1y Lys Lys Thr G1n Arg Ser Leu Ser Ala Glu Leu Glu Ile Pro Ala Val Lys Glu Ser Asp Ala Gly Lys Tyr Tyr Cys Arg Ala Asp Asn Gly His Val Pro Ile Gln Ser Lys Val Val Asn Ile Pro Val Arg Ile Pro Val Ser Arg Pro Val Leu Thr Leu Arg Ser Pro Gly Ala Gln Ala Ala Val Gly Asp Leu Leu Glu Leu His Cys Glu Ala Leu Arg Gly Ser Pro Pro Tle Leu Tyr Gln Phe Tyr His Glu Asp Val Thr Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Ala Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asn Asn G1y Leu Gly Ala Gln Cys Ser Glu Ala Val Pro Val Ser Ile Ser Gly Pro Asp Gly Tyr Arg Arg Asp Leu Met Thr Ala Gly Val Leu Trp Gly Leu Phe Gly Val Leu Gly Phe Thr Gly Val Ala Leu Leu Leu Tyr Ala Leu Phe His Lys Tle Ser Gly Glu Ser Ser Ala Thr Asn Glu Pro Arg G1y Ala Ser Arg Pro Asn Pro Gln Glu Phe Thr Tyr Ser Ser Pro Thr Pro Asp Met Glu Glu Leu Gln Pro Val Tyr Val Asn Val Gly Ser Val Asp Val Asp Val Val Tyr Ser Gln Val Trp Ser Met Gln Gln Pro Glu Ser Ser Ala Asn Ile Arg Thr Leu Leu Glu Asn Lys Asp Ser Gln Val 21e Tyr Ser Ser Val Lys Lys Ser <210> 5 <211> 140 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 5 His Tyr Ala Arg Ala Arg Arg Lys Pro Gly Gly Leu Ser Ala Thr Gly Thr Ser Ser His Ser Pro Ser Glu Cys Gln Glu Pro Ser Ser Ser Arg Pro Ser Arg Ile Asp Pro Gln G1u Pro Thr His Ser Lys Pro Leu Ala Pro Met Glu Leu Glu Pro Met Tyr Ser Asn Val Asn Pro Gly Asp Ser Asn Pro Ile Tyr Ser Gln Ile Trp Ser Ile Gln His Thr Lys Glu Asn Ser Ala Asn Cys Pro Met Met His Gln Glu His Glu Glu Leu Thr Val Leu Tyr Ser Glu Leu Lys Lys Thr His Pro Asp Asp Ser Ala Gly Glu Ala Ser Ser Arg Gly Arg Ala His Glu Glu Asp Asp Glu Glu Asn Tyr Glu Asn Val Pro Arg Val Leu Leu Ala Ser Asp His <210> 6 <211> 717 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 6 Gly Val Ala Pro Lys Ala Val Leu Leu Leu Asn Pro Pro Trp Ser Thr Ala Phe Lys Gly Glu Lys Val Ala Leu Ile Cys Ser Ser Ile Ser His Ser Leu Ala Gln Gly Asp Thr Tyr Trp Tyr His Asp Glu Lys Leu Leu Lys Ile Lys His Asp Lys Ile Gln Ile Thr Glu Pro Gly Asn Tyr Gln Cys Lys Thr Arg Gly Ser Ser Leu Ser Asp Ala Val His Val Glu Phe Ser Pro Asp Trp Leu Ile Leu Gln Ala Leu His Pro Val Phe Glu Gly Asp Asn Val Ile Leu Arg Cys Gln Gly Lys Asp Asn Lys Asn Thr His Gln Lys Val Tyr Tyr Lys Asp Gly Lys Gln Leu Pro Asn Ser Tyr Asn 1l5 120 125 Leu Glu Lys Ile Thr Val Asn Ser Val Ser Arg Asp Asn Ser Lys Tyr His Cys Thr Ala Tyr Arg Lys Phe Tyr Ile Leu Asp Ile Glu Val Thr Ser Lys Pro Leu Asn Ile Gln Val Gln Glu Leu Phe Leu His Pro Val Leu Arg Ala Ser Ser Ser Thr Pro Ile Glu Gly Ser Pro Met Thr Leu Thr Cys Glu Thr Gln Leu Ser Pro Gln Arg Pro Asp Val Gln Leu Gln Phe Ser Leu Phe Arg Asp Ser Gln Thr Leu Gly Leu Gly Trp Ser Arg Ser Pro Arg Leu Gln Ile Pro Ala Met Trp Thr Glu Asp Ser Gly Ser Tyr Trp Cys G1u Va1 Glu Thr Va1 Thr His Ser Ile Lys Lys Arg Ser Leu Arg Ser Gln Ile Arg Val Gln Arg Val Pro Val Ser Asn Val Asn Leu Glu Ile Arg Pro Thr Gly Gly Gln Leu Ile Glu Gly Glu Asn Met Val Leu Ile Cys Ser Val Ala Gln Gly Ser Gly Thr Val Thr Phe Ser Trp His Lys Glu Gly Arg Val Arg Ser Leu Gly Arg Lys Thr G1n Arg Ser Leu Leu Ala G1u Leu His Val Leu Thr Val Lys Glu Ser Asp Ala Gly Arg Tyr Tyr Cys Ala Ala Asp Asn Val His Ser Pro Ile Leu Ser Thr Trp Ile Arg Val Thr Val Arg Tle Pro Val Ser His Pro Val Leu Thr Phe Arg Ala Pro Arg Ala His Thr Val Val Gly Asp Leu Leu Glu Leu His Cys Glu Ser Leu Arg Gly Ser Pro Pro Ile Leu Tyr Arg Phe Tyr His Glu Asp Val Thr Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Ala Glu His Ser Gly Asn Tyr Ser Cys Asp Ala Asp Asn Gly Leu Gly A1a Gln His Ser His Gly Val Ser Leu Arg Val Thr Val Pro Val Ser Arg Pro Val Leu Thr Leu Arg Ala Pro Gly Ala Gln Ala Val Val Gly Asp Leu Leu Glu Leu His Cys Glu Ser Leu Arg Gly Ser Phe Pro Ile Leu Tyr Trp Phe Tyr His Glu Asp Asp Thr Leu Gly Asn Ile Ser Ala His Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Thr Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asp Asn Gly Leu Gly Ala G1n His Ser Lys Val Val Thr Leu Asn Val Thr Gly Thr Ser Arg Asn Arg Thr Gly Leu Thr Ala Ala Gly Ile Thr Gly Leu Val Leu Ser Ile Leu Val Leu Ala Ala Ala Ala Ala Leu Leu His Tyr Ala Arg Ala Arg Arg Lys Pro Gly Gly Leu 5er A1a Thr Gly Thr Ser Ser His Ser Pro Ser Glu Cys Gln Glu Pro Ser Ser Ser Arg Pro Ser Arg Ile Asp Pro Gln Glu Pro Thr His Ser Lys Pro Leu Ala Pro Met Glu Leu Glu Pro Met Tyr Ser Asn Val Asn Pro Gly Asp Ser Asn Pro Ile Tyr Ser Gln Ile Trp Ser Ile Gln His Thr Lys Glu Asn Ser Ala Asn Cys Pro Met Met His Gln Glu His Glu Glu Leu Thr Val Leu Tyr Ser Glu Leu Lys Lys Thr His Pro Asp Asp Ser Ala Gly Glu Ala Ser Ser Arg Gly Arg Ala His Glu Glu Asp Asp Glu Glu Asn Tyr Glu Asn Val Pro Arg Val Leu Leu Ala Ser Asp His <210> 7 <211> 300 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 7 aaaaa taggaagacg ttcagccagggatccactcaggagccttcccagccctcta 60 aaaa g accccagggcagctacagcctatatatgaa 120 ccccaagagt tcacctacct caactcacct aatgtgaatg ttgtaagtgg ggatgaggtttattcactggcgtactataaccagccggag 180 caggaatcag tagcagcaga aaccctggggacacatatggaggacaaggtttccttagac 240 atctattcca ggctgaggaa agcaaacattacagatgtggactatgaagatgctatgtaa 300 <210> 8 <211> 2038 <212> DNA

<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 8 ctcgactctg aggtgcattc tttttttgat gagaggcatc tctaggtacc atccctgacc 60 tggtcctcat gctgccgagg ctgttgctgt tgatctgtgc tccactctgt gaacctgccg 120 agctgtttttgatagccagcccctcccatcccacagaggggagcccagtgaccctgacgt180 gtaagatgccctttctacagagttcagatgcccagttccagttctgctttttcagagaca240 cccgggccttgggcccaggctggagcagctcccccaagctccagatcgctgccatgtgga300 aagaagacacagggtcatactggtgcgaggcacagacaatggcgtccaaagtcttgagga360 gcaggagatcccagataaatgtgcacagggtccctgtcgctgatgtgagcttggagactc420 agcccccaggaggacaggtgatggagggagacaggctggtcctcatctgctcagttgcta480 tgggcacaggagacatcaccttcctttggtacaaaggggctgtaggtttaaaccttcagt540 caaagacccagcgttcactgacagcagagtatgagattccttcagtgagggagagtgatg600 ctgagcaatattactgtgtagctgaaaatggctatggtcccagccccagtgggctggtga660 gcatcactgtcagaatcccggtgtctcgcccaatcctcatgctcagggctcccagggccc720 aggctgcagtggaggatgtgctggagcttcactgtgaggccctgagaggctctcctccaa780 tcctgtactggttttatcacgaggatatcaccctggggagcaggtcggccccctctggag840 gaggagcctccttcaacctttccctgactgaagaacattctggaaactactcctgtgagg900 ccaacaatggcctgggggcccagcgcagtgaggcggtgacactcaacttcacagtgccta960 ctggggccagaagcaatcatcttacctcaggagtcattgaggggctgctcagcacccttg1020 gtccagccaccgtggccttattattttgctacggcctcaaaagaaaaataggaagacgtt1080 cagccagggatccactcaggagccttcccagccctctaccccaagagttcacctacctca1140 actcacctaccccagggcagctacagcctatatatgaaaatgtgaatgttgtaagtgggg1200 atgaggtttattcactggcgtactataaccagccggagcaggaatcagtagcagcagaaa1260 ccctggggacacatatggaggacaaggtttccttagacatctattccaggctgaggaaag1320 caaacattacagatgtggactatgaagatgctatgtaaggttatggaagattctgctctt1380 tgaaaaccatccatgaccccaagcctcaggcctgatatgttcttcagagatcctggggca1440 ttagctttccagtatacctcttctggatgccattctccatggcactattccttcatctac1500 tgtgaagtgaagttggcgcagccctgaagaaactacctaggagaactaatagacacagga1560 gtgacagggactttgttatcagaaccagattcctgccggctcctttgaaaacaggtcata1620 ttgtgctcttctgtttacaagaggaaacaagatggaataaaagaaattgggatcttgggt1680 tggagggacagtgaagcttagagcacatgaactcaaggttagtgactctgcaggacttca1740 cagagagagctgtgcccatcattcagtccaagtgctttctctgcccagacagcacagaac1800 tccagccccgctacttacatggatcatcgagtttccacctaaaatatgattctatttatt1860 ttgagtcactgttaccaaattagaactaaaacaaagttacataaaaagttattgtgactc1920 cacttaattttagtgacgtatttttgtatatataggccaacctataccacatccaaaatt1980 atgtatctattacagcccctagaagctttataaatacagtgtgtcttcttttattcac 2038 <210> 9 <211> 261 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificialequence:/note =
S

synthetic construct <400> 9 cacaagatat caggagaaag ttctgccactaatgaacccagaggggcttccaggccaaat60 cctcaagagt tcacctattc aagcccaaccccagacatggaggagctgcagccagtgtat120 gtcaatgtgg gctctgtaga tgtggatgtggtttattctcaggtctggagcatgcagcag180 ccagaaagct cagcaaacat caggacacttctggagaacaaggactcccaagtcatctac240 tcttctgtga agaaatcata a 261 <210> 10 <211> 2573 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 10 ggtgaccaag agtacatctc ttttcaaatagctggattaggtcctcatgctgctgtggtc60 attgctggtc atctttgatg cagtcactgaacaggcagattcgctgacccttgtggcgcc120 ctcttctgtcttcgaaggagacagcatcgttctgaaatgccagggagaacagaactggaa180 aattcagaagatggcttaccataaggataacaaagagttatctgttttcaaaaaattctc240 agatttccttatccaaagtgcagttttaagtgacagtggtaactatttctgtagtaccaa300 aggacaactctttctctgggataaaacttcaaatatagtaaagataaaagtccaagagct360 ctttcaacgtcctgtgctgactgccagctccttccagcccatcgaagggggtccagtgag420 cctgaaatgtgagacccggctctctccacagaggttggatgttcaactccagttctgctt480 cttcagagaaaaccaggtcctggggtcaggctggagcagctctccggagctccagatttc540 tgccgtgtggagtgaagacacagggtcttactggtgcaaggcagaaacggtgactcacag600 gatcagaaaacagagcctccaatcccagattcacgtgcagagaatccccatctctaatgt660 aagcttggagatccgggcccccgggggacaggtgactgaaggacaaaaactgatcctgct720 ctgctcagtggctgggggtacaggaaatgtcacattctcctggtacagagaggccacagg780 aaccagtatgggaaagaaaacccagcgttccctgtcagcagagctggagatcccagctgt840 gaaagagagtgatgccggcaaatattactgtagagctgacaacggccatgtgcctatcca900 gagcaaggtggtgaatatccctgtgagaattccagtgtctcgccctgtcctcaccctcag960 gtctcctggggcccaggctgcagtgggggacctgctggagcttcactgtgaggccctgag1020 aggctctcccccaatcttgtaccaattttatcatgaggatgtcacccttgggaacagctc1080 ggccccctctggaggaggggcctccttcaacctctctttgactgcagaacattctggaaa1140 ctactcctgtgaggccaacaacggcctgggggcccagtgcagtgaggcagtgccagtctc1200 catctcaggacctgatggctatagaagagacctcatgacagctggagttctctggggact1260 gtttggtgtccttggtttcactggtgttgctttgctgttgtatgccttgttccacaagat1320 atcaggagaaagttctgccactaatgaacccagaggggcttccaggccaaatcctcaaga1380 gttcacctattcaagcccaaccccagacatggaggagctgcagccagtgtatgtcaatgt1440 gggctctgtagatgtggatgtggtttattctcaggtctggagcatgcagcagccagaaag1500 ctcagcaaacatcaggacacttctggagaacaaggactcccaagtcatctactcttctgt1560 gaagaaatcataacacttggaggaatcagaagggaagatcaacagcaaggatggggcatc1620 attaagacttgctataaaaccttatgaaaatgcttgaggcttatcacctgccacagccag1680 aacgtgcctcaggaggcacctcctgtcatttttgtcctgatgatgtttcttctccaatat1740 cttcttttacctatcaatattcattgaactgctgctacatccagacactgtgcaaataaa1800 ttatttctgctaccttctcttaagcaatcagtgtgtaaagatttgagggaagaatgaata1860 agagatacaaggtctcaccttcatctactgtgaagtgatgagaacaggacttgatagtgg1920 tgtattaacttatttatgtgctgctggatacagtttgctaatattttgttgagaattttt1980 gcaaatatgttcattgggaatattggcctgaaattttcttttccactgtgtctctgccag2040 aatgtttgtatcaggctgatgctggcttcatagaatgagttaggcaggagcccttcctcc2100 ttgattttttggcatagtttcagcaggattggtaccagttattctttctgcatcttgtag2160 aattcagctatgaatccatctggtctagggcttttgtgttggttggtaagttttttatta2220 ctaattcaacttcagcgcttgatattggtctaggaggggtttctgtctcttcctggttca2280 atcttgggagattgtgtgtttccaggaatttagccgtttcctccagattttcttctttat2340 gtgcatcgacttgagtgtaaacataacttatatgcactgggaaaccaaaaaatctgtgtg2400 acttgctttattgcagcatttgttttattttggtagtctggaactgaacctgcaatatca2460 ccaaagtatgcatatagttgcaaaaatgtgatttttgacatagtaaatatgagtatttgc2520 aataaactatgatattacttttgtaagtatatagaataaaatgtaaataatct 2573 <210> 11 <211> 423 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
11 gggcccgaaggaaaccaggaggactttctgccactggaacatctagtcac 60 cattacgcca agtcctagcgagtgtcaggagccttcctcgtccaggccttccaggatagaccctcaagag 120 cccactcactctaaaccactagccccaatggagctggagccaatgtacagcaatgtaaat 180 cctggagatagcaacccgatttattcccagatctggagcatccagcatacaaaagaaaac 240 tcagctaattgtccaatgatgcatcaagagcatgaggaacttacagtcctctattcagaa 300 ctgaagaagacacacccagacgactctgcaggggaggctagcagcagaggcagggcccat 360 gaagaagatgatgaagaaaactatgagaatgtaccacgtgtattactggcctcagaccac 420 tag 423 <210> 12 <211> 2916 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
12 agtagcagcttcctgccctccttcttggagataagtcgggcttttggtga60 gtctcatctg gacagactttcccaaccctctgcccggccggtgcccatgcttctgtggctgctgctgctg120 atcctgactcctggaagagaacaatcaggggtggccccaaaagctgtacttctcctcaat180 cctccatggtccacagccttcaaaggagaaaaagtggctctcatatgcagcagcatatca240 cattccctagcccagggagacacatattggtatcacgatgagaagttgttgaaaataaaa300 catgacaagatccaaattacagagcctggaaattaccaatgtaagacccgaggatcctcc360 ctcagtgatgccgtgcatgtggaattttcacccgactggctgatcctgcaggctttacat420 cctgtctttgaaggagacaatgtcattctgagatgtcaggggaaagacaacaaaaacact480 catcaaaaggtttactacaaggatggaaaacagcttcctaatagttataatttagagaag540 atcacagtgaattcagtctccagggataatagcaaatatcattgtactgcttataggaag600 ttttacatacttgacattgaagtaacttcaaaacccctaaatatccaagttcaagagctg660 tttctacatcctgtgctgagagccagctcttccacgcccatagaggggagtcccatgacc720 ctgacctgtgagacccagctctctccacagaggccagatgtccagctgcaattctccctc780 ttcagagatagccagaccctcggattgggctggagcaggtcccccagactccagatccct840 gccatgtggactgaagactcagggtcttactggtgtgaggtggagacagtgactcacagc900 atcaaaaaaaggagcctgagatctcagatacgtgtacagagagtccctgtgtctaatgtg960 aatctagagatccggcccaccggagggcagctgattgaaggagaaaatatggtccttatt1020 tgctcagtagcccagggttcagggactgtcacattctcctggcacaaagaaggaagagta1080 agaagcctgggtagaaagacccagcgttccctgttggcagagctgcatgttctcaccgtg1140 aaggagagtgatgcagggagatactactgtgcagctgataacgttcacagccccatcctc1200 agcacgtggattcgagtcaccgtgagaattccggtatctcaccctgtcctcaccttcagg gctcccagggcccacactgtggtgggggacctgctggagcttcactgtgagtccctgaga1320 ggctctcccccgatcctgtaccgattttatcatgaggacgtcaccctggggaacagctca1380 gccccctctggaggaggagcctccttcaacctctctctgactgcagaacattctggaaac1440 tactcctgtgatgcagacaatggcctgggggcccagcacagtcatggagtgagtctcagg1500 gtcacagttccggtgtctcgccccgtcctcaccctcagggctcccggggcccaggctgtg1560 gtgggggacctgctggagcttcactgtgagtccctgagaggctccttcccgatcctgtac1620 tggttttatcacgaggatgacaccttggggaacatctcggcccactctggaggaggggca1680 tccttcaacctctctctgactacagaacattctggaaactactcatgtgaggctgacaat1740 ggcctgggggcccagcacagtaaagtggtgacactcaatgttacaggaacttccaggaac1800 agaacaggccttaccgctgcgggaatcacggggctggtgctcagcatcctcgtccttgct1860 gctgctgctgctctgctgcattacgccagggcccgaaggaaaccaggaggactttctgcc1920 actggaacatctagtcacagtcctagcgagtgtcaggagccttcctcgtccaggccttcc1980 aggatagaccctcaagagcccactcactctaaaccactagccccaatggagctggagcca2040 atgtacagcaatgcaaatcctggagatagcaacccgatttattcccagatctggagcatc2100 cagcatacaaaagaaaactcagctaattgtccaatgatgcatcaagagcatgaggaactt2160 acagtcctctattcagaactgaagaagacacacccagacgactctgcaggggaggctagc2220 agcagaggcagggcccatgaagaagatgatgaagaaaactatgagaatgtaccacgtgta2280 ttactggcctcagaccactagccccttacccagagtggcccacaggaaacagcctgcacc2340 atttttttttctgttctctccaaccacacatcatccatctctccagactctgcctcctac2400 gaggctgggctgcagg 2416 <210> 13 <211> 873 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
13 tgatagccagcccctcccatcccacagaggggagcccagtgaccctgacg 60 gagctgtttt tgtaagatgccctttctacagagttcagatgcccagttccagttctgctttttcagagac 120 acccgggccttgggcccaggctggagcagctcccccaagctccagatcgctgccatgtgg 180 aaagaagacacagggtcatactggtgcgaggcacagacaatggcgtccaaagtcttgagg 240 agcaggagatcccagataaatgtgcacagggtccctgtcgctgatgtgagcttggagact 300 cagcccccaggaggacaggtgatggagggagacaggctggtcctcatctgctcagttgct 360 atgggcacaggagacatcaccttcctttggtacaaaggggctgtaggtttaaaccttcag 420 tcaaagacccagcgttcactgacagcagagtatgagattccttcagtgagggagagtgat 480 gctgagcaatattactgtgtagctgaaaatggctatggtcccagccccagtgggctggtg 540 agcatcactgtcagaatcccggtgtctcgcccaatcctcatgctcagggctcccagggcc 600 caggctgcagtggaggatgtgctggagcttcactgtgaggccctgagaggctctcctcca 660 atcctgtactggttttatcacgaggatatcaccctggggagcaggtcggccccctctgga 720 ggaggagcctccttcaacctttccctgactgaagaacattctggaaactactcctgtgag 780 gccaacaatggcctgggggcccagcgcagtgaggcggtgacactcaacttcacagtgcct 840 actggggccagaagcaatcatcttacctcagga 873 <210>

<211>

<212>
DNA

<213> ficial Arti Sequence <220>

<223> iption equence:/note =
Descr of Artificial S

synth etic construct <400>
14 cgccctcttctgtcttcgaaggagacagcatcgttctgaaatgccaggga60 acccttgtgg gaacagaactggaaaattcagaagatggcttaccataaggataacaaagagttatctgtt120 ttcaaaaaattctcagatttccttatccaaagtgcagttttaagtgacagtggtaactat180 ttctgtagtaccaaaggacaactctttctctgggataaaacttcaaatatagtaaagata290 aaagtccaagagctctttcaacgtcctgtgctgactgccagctccttccagcccatcgaa300 gggggtccagtgagcctgaaatgtgagacccggctctctccacagaggttggatgttcaa360 ctccagttctgcttcttcagagaaaaccaggtcctggggtcaggctggagcagctctccg420 gagctccagatttctgccgtgtggagtgaagacacagggtcttactggtgcaaggcagaa480 acggtgactcacaggatcagaaaacagagcctccaatcccagattcacgtgcagagaatc540 cccatctctaatgtaagcttggagatccgggcccccgggggacaggtgactgaaggacaa600 , aaactgatcctgctctgctcagtggctgggggtacaggaaatgtcacattctcctggtac660 agagaggccacaggaaccagtatgggaaagaaaacccagcgttccctgtcagcagagctg720 gagatcccagctgtgaaagagagtgatgccggcaaatattactgtagagctgacaacggc780 catgtgcctatccagagcaaggtggtgaatatccctgtgagaattccagtgtctcgccct840 gtcctcaccctcaggtctcctggggcccaggctgcagtgggggacctgctggagcttcac900 tgtgaggccctgagaggctctcccccaatcttgtaccaattttatcatgaggatgtcacc960 cttgggaacagctcggccccctctggaggaggggcctccttcaacctctctttgactgca1020 gaacattctggaaactactcctgtgaggccaacaacggcctgggggcccagtgcagtgag1080 gcagtgccagtctccatctcaggacctgatggctatagaagagacctcatgacagct 1137 <210>

<211>

<212>
DNA

<213> ence Artificial Sequ <220>
<223> Description of Artificial Sequence:/note synthetic construct <400> 15 gtggccccaa aagctgtact tctcctcaat cctccatggt ccacagcctt caaaggagaa 60 aaagtggctc tcatatgcag cagcatatca cattccctag cccagggaga cacatattgg 120 tatcacgatg agaagttgtt gaaaataaaa catgacaaga tccaaattac agagcctgga 180 aattaccaat gtaagacccg aggatcctcc ctcagtgatg ccgtgcatgt ggaattttca 240 cccgactggctgatcctgcaggctttacatcctgtctttgaaggagacaatgtcattctg300 agatgtcaggggaaagacaacaaaaacactcatcaaaaggtttactacaaggatggaaaa360 cagcttcctaatagttataatttagagaagatcacagtgaattcagtctccagggataat420 agcaaatatcattgtactgcttataggaagttttacatacttgacattgaagtaacttca480 aaacccctaaatatccaagttcaagagctgtttctacatcctgtgctgagagccagctct540 tccacgcccatagaggggagtcccatgaccctgacctgtgagacccagctctctccacag600 aggccagatgtccagctgcaattctccctcttcagagatagccagaccctcggattgggc660 tggagcaggtcccccagaotccagatccctgccatgtggactgaagactcagggtcttac720 tggtgtgaggtggagacagtgactcacagcatcaaaaaaaggagcctgagatctcagata780 cgtgtacagagagtccctgtgtctaatgtgaatctagagatccggcccaccggagggcag840 ctgattgaaggagaaaatatggtccttatttgctcagtagcccagggttcagggactgtc900 acattctcctggcacaaagaaggaagagtaagaagcctgggtagaaagacccagcgttcc960 ctgttggcagagctgcatgttctcaccgtgaaggagagtgatgcagggagatactactgt1020 gcagctgataacgttcacagccccatcctcagcacgtggattcgagtcaccgtgagaatt1080 ccggtatctcaccctgtcctcaccttcagggctcccagggcccacactgtggtgggggac1140 ctgctggagcttcactgtgagtccctgagaggctctcccccgatcctgtaccgattttat1200 catgaggacgtcaccctggggaacagctcagccccctctggaggaggagcctccttcaac1260 ctctctctgactgcagaacattctggaaactactcctgtgatgcagacaatggcctgggg1320 gcccagcacagtcatggagtgagtctcagggtcacagttccggtgtctcgccccgtcctc1380 accctcagggctcccggggcccaggctgtggtgggggacctgctggagcttcactgtgag1440 tccctgagaggctccttcccgatcctgtactggttttatcacgaggatgacaccttgggg1500 aacatctcggcccactctggaggaggggcatccttcaacctctctctgactacagaacat1560 tctggaaactactcatgtgaggctgacaatggcctgggggcccagcacagtaaagtggtg1620 acactcaatgttacaggaacttccaggaacagaacaggc 1659 <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>

<223> iption rtificialequence:/note =
Descr of A S

synth etic construct <400>
16 gggcccgaaggaaaccaggaggactttctgccactggaacatctagtcac60 cattacgcca agtcctagcgagtgtcaggagccttcctcgtccaggccttccaggatagaccctcaagag120 cccactcactctaaaccactagccccaatggagctggagccaatgtacagcaatgcaaat180 cctggagatagcaacccgatttattcccagatctggagcatccagcatacaaaagaaaac240 tcagctaattgtccaatgatgcatcaagagcatgaggaacttacagtcctctattcagaa300 ctgaagaagacacacccagacgactctgcaggggaggctagcagcagaggcagggcccat360 gaagaagatgatgaagaaaactatgagaatgtaccacgtgtattactggcctcagaccac420 tag <210>

<211>

<212>
DNA

<213>
Artificial Sequence <220>

<223>
Description of Artificial Sequence:/note =

synthetic construct <400>
17 aagctgtacttctcctcaatcctccatggtccacagccttcaaaggagaa60 gtggccccaa aaagtggctctcatatgcagcagcatatcacattccctagcccagggagacacatattgg120 tatcacgatgagaagttgttgaaaataaaacatgacaagatccaaattacagagcctgga180 aattaccaatgtaagacccgaggatcctccctcagtgatgccgtgcatgtggaattttca240 cccgactggctgatcctgcaggctttacatcctgtctttgaaggagacaatgtcattctg300 agatgtcaggggaaagacaacaaaaacactcatcaaaaggtttactacaaggatggaaaa360 cagcttcctaatagttataatttagagaagatcacagtgaattcagtctccagggataat420 agcaaatatcattgtactgcttataggaagttttacatacttgacattgaagtaacttca480 aaacccctaaatatccaagttcaagagctgtttctacatcctgtgctgagagccagctct540 tccacgcccatagaggggagtcccatgaccctgacctgtgagacccagctctctccacag600 aggccagatgtccagctgcaattctccctcttcagagatagccagaccctcggattgggc660 tggagcaggtcccccagactccagatccctgccatgtggactgaagactcagggtcttac720 tggtgtgaggtggagacagtgactcacagcatcaaaaaaaggagcctgagatctcagata780 cgtgtacagagagtccctgtgtctaatgtgaatctagagatccggcccaccggagggcag840 ctgattgaaggagaaaatatggtccttatttgctcagtagcccagggttcagggactgtc900 acattctcctggcacaaagaaggaagagtaagaagcctgggtagaaagacccagcgttcc960 ctgttggcagagctgcatgttctcaccgtgaaggagagtgatgcagggagatactactgt1020 gcagctgataacgttcacagccccatcctcagcacgtggattcgagtcaccgtgagaatt1080 ccggtatctcaccctgtcctcaccttcagggctcccagggcccacactgtggtgggggac1140 ctgctggagcttcactgtgagtccctgagaggctctcccccgatcctgtaccgattttat1200 catgaggacgtcaccctggggaacagctcagccccctctggaggaggagcctccttcaac1260 ctctctctgactgcagaacattctggaaactactcctgtgatgcagacaatggcctgggg1320 gcccagcacagtcatggagtgagtctcagggtcacagttccggtgtctcgccccgtcctc1380 accctcagggctcccggggcccaggctgtggtgggggacctgctggagcttcactgtgag1440 tccctgagaggctccttcccgatcctgtactggttttatcacgaggatgacaccttgggg1500 aacatctcggcccactctggaggaggggcatccttcaacctctctctgactacagaacat1560 tctggaaactactcatgtgaggctgacaatggcctgggggcccagcacagtaaagtggtg1620 acactcaatgttacaggaacttccaggaacagaacaggccttaccgctgcgggaatcacg1680 gggctggtgctcagcatcctcgtccttgctgctgctgctgctctgctgcattacgccagg1740 gcccgaaggaaaccaggaggactttctgccactggaacatctagtcacagtcctagcgag1800 tgtcaggagccttcctcgtccaggccttccaggatagaccctcaagagcccactcactct1860 aaaccactagccccaatggagctggagccaatgtacagcaatgcaaatcctggagatagc1920 aacccgatttattcccagatctggagcatccagcatacaaaagaaaactcagctaattgt1980 ccaatgatgcatcaagagcatgaggaacttacagtcctctattcagaactgaagaagaca2040 cacccagacgactctgcaggggaggctagcagcagaggcagggcccatgaagaagatgat2100 gaagaaaactatgagaatgtaccacgtgtattactggcctcagaccactag 2151 <210> 18 <211> 315 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificialequence:/note =
S

synthetic construct <400> 18 agatcctgga gaaaagctgg gccccttccatcccagataccacccacagctccaggtgga60 gagcagtgcc cactatatgc caacgtgcatcaccagaaagggaaagatgaaggtgttgtc120 tactctgtgg tgcatagaac ctcaaagaggagtgaagccaggtctgctgagttcaccgtg180 gggagaaagg acagttctat catctgtgcggaggtgagatgcctgcagcccagtgaggtt240 tcatccacgg aggtgaatat gagaagcaggactctccaagaaccccttagcgactgtgag300 gaggttctct gctag 315 <210> 19 <211> 870 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 19 actgtctggc tgtacctcca agcctggccaaaccctgtgtttgaaggagatgccctgact60 ctgcgatgtc agggatggaa gaatacaccactgtctcaggtgaagttctacagagatgga120 aaattccttc atttctctaa ggaaaaccagactctgtccatgggagcagcaacagtgcag180 agccgtggcc agtacagctg ctctgggcaggtgatgtatattccacagacattcacacaa240 acttcagagactgccatggttcaagtccaagagctgtttccacctcctgtgctgagtgcc300 atcccctctcctgagccccgagagggtagcctggtgaccctgagatgtcagacaaagctg360 caccccctgaggtcagccttgaggctccttttctccttccacaaggacggccacaccttg920 caggacaggggccctcacccagaactctgcatcccgggagccaaggagggagactctggg480 ctttactggtgtgaggtggcccctgagggtggccaggtccagaagcagagcccccagctg590 gaggtcagagtgcaggctcctgtatcccgtcctgtgctcactctgcaccacgggcctgct600 gaccctgctgtgggggacatggtgcagctcctctgtgaggcacagaggggctcccctccg660 atcctgtattccttctaccttgatgagaagattgtggggaaccactcagctccctgtggt720 ggaaccacctccctcctcttcccagtgaagtcagaacaggatgctgggaactactcctgc780 gaggctgagaacagtgtctccagagagaggagtgagcccaagaagctgtctctgaagggt840 tctcaagtcttgttcactcccgccagcaac 870 <210>

<211>

<212>
DNA

<213> ficial enoe Arti Sequ <220>
<223> Description of Artificial Sequence:/note synthetic construct <400>
20 tgtacctccaagcctggccaaaccctgtgtttgaaggagatgccctgact60 actgtctggc ctgcgatgtcagggatggaagaatacaccactgtctcaggtgaagttctacagagatgga120 aaattccttcatttctctaaggaaaaccagactctgtccatgggagcagcaacagtgcag180 agccgtggccagtacagctgctctgggcaggtgatgtatattccacagacattcacacaa240 acttcagagactgccatggttcaagtccaagagctgtttccacctcctgtgctgagtgcc300 atcccctctcctgagccccgagagggtagcctggtgaccctgagatgtcagacaaagctg360 caccccctgaggtcagccttgaggctccttttctccttccacaaggacggccacaccttg420 caggacaggggccctcacccagaactctgcatcccgggagccaaggagggagactctggg480 ctttactggtgtgaggtggcccctgagggtggccaggtccagaagcagagcccccagctg540 gaggtcagagtgcaggctcctgtatcccgtcctgtgctcactctgcaccacgggcctgct600 gaccctgctgtgggggacatggtgcagctcctctgtgaggcacagaggggctcccctccg660 atcctgtattccttctaccttgatgagaagattgtggggaaccactcagctccctgtggt720 ggaaccacctccctcctcttcccagtgaagtcagaacaggatgctgggaactactcctgc780 gaggctgagaacagtgtctccagagagaggagtgagcccaagaagctgtctctgaagggt840 tctcaagtcttgttcactcccgccagcaactggctggttccttggcttcctgcgagcctg900 cttggcctgatggttattgctgctgcacttctggtttatgtgagatcctggagaaaagct960 gggccccttccatcccagataccacccacagctccaggtggagagcagtgcccactatat1020 gccaacgtgcatcaccagaaagggaaagatgaaggtgttgtctactctgtggtgcataga1080 acctcaaagaggagtgaagccaggtctgctgagttcaccgtggggagaaaggacagttct1140 atcatctgtgcggaggtgagatgcctgcagcccagtgaggtttcatccacggaggtgaat1200 atgagaagcaggactctccaagaaccccttagcgactgtgaggaggttctctgctag 1257 <210>

<211>

<212>
PRT

<213> ficial ence Arti Sequ <220>

<223> ription ArtificialSequence:/note =
Desc of synt hetic construct <400>
21 Pro Ser Pro Thr Ala Glu His Glu Gly Leu Phe Ser Leu Ile Ala Ser Pro Va1 Pro Phe Thr Leu Leu Thr Cys Gln Lys Met Ser Ser Asp Ala Gln Phe Asp Thr Gln Phe Arg Cys Phe Ala Phe Arg Leu Gly Pro Gly Trp Ser Ser Ser Pro Lys Leu Gln Ile Ala Ala Met Trp Lys Glu Asp Thr Gly Ser Tyr Trp Cys Glu Ala Gln Thr Met Ala Ser Lys Val Leu Arg Ser Arg Arg Ser Gln Ile Asn Val His Arg Val Pro Val Ala Asp Val Ser Leu Glu Thr Gln Pro Pro Gly Gly G1n Val Met Glu Gly Asp Arg Leu Val Leu Ile Cys Ser Val Ala Met Gly Thr Gly Asp Ile Thr Phe Leu Trp Tyr Lys Gly Ala Val Gly Leu Asn Leu Gln Ser Lys Thr Gln Arg Ser Leu Thr Ala Glu Tyr Glu Ile Pro Ser Val Arg Glu Ser Asp A1a Glu Gln Tyr Tyr Cys Val Ala Glu Asn Gly Tyr Gly Pro Ser Pro Ser Gly Leu Val Ser Ile Thr Val Arg Ile Pro Val Ser Arg Pro Ile Leu Met Leu Arg Ala Pro Arg Ala Gln Ala Ala Val Glu Asp Val Leu Glu Leu His Cys Glu Ala Leu Arg Gly Ser Pro Pro Ile Leu Tyr Trp Phe Tyr His Glu Asp Ile Thr Leu Gly Ser Arg Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Glu Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asn Asn Gly Leu Gly Ala Gln Arg Ser Glu Ala Val Thr Leu Asn Phe Thr Val Pro Thr Gly Ala Arg Ser Asn His Leu Thr Ser Gly <210> 22 <211> 380 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial 5equence:/note =
synthetic construct <400> 22 Leu Thr Leu Val Ala Pro Ser Ser Val Phe Glu Gly Asp Ser Ile Val Leu Lys Cys Gln Gly Glu Gln Asn Trp Lys Ile Gln Lys Met Ala Tyr His Lys Asp Asn Lys Glu Leu Ser Val Phe Lys Lys Phe Ser Asp Phe Leu Ile Gln Ser Ala Val Leu Ser Asp Ser G1y Asn Tyr Phe Cys Ser Thr Lys Gly Gln Leu Phe Leu Trp Asp Lys Thr Ser Asn Ile Val Lys Tle Lys Val Gln Glu Leu Phe Gln Arg Pro Val Leu Thr Ala Ser Ser Phe Gln Pro Ile Glu Gly Gly Pro Val Ser Leu Lys Cys Glu Thr Arg Leu Ser Pro Gln Arg Leu Asp Val Gln Leu Gln Phe Cys Phe Phe Arg Glu Asn Gln Val Leu Gly Ser Gly Trp Ser Ser Ser Pro Glu Leu Gln Ile Ser Ala Val Trp Ser Glu Asp Thr Gly Ser Tyr Trp Cys Lys Ala Glu Thr Val Thr His Arg Ile Arg Lys Gln Ser Leu Gln Ser Gln Ile His Val Gln Arg Ile Pro Ile Ser Asn Val Ser Leu Glu Ile Arg Ala Pro Gly Gly Gln Val Thr Glu Gly Gln Lys Leu Ile Leu Leu Cys Ser Val Ala Gly Gly Thr Gly Asn Val Thr Phe Ser Trp Tyr Arg Glu Ala Thr G1y Thr Ser Met Gly Lys Lys Thr Gln Arg Ser Leu Ser Ala Glu Leu Glu Ile Pro Ala Val Lys Glu Ser Asp Ala Gly Lys Tyr Tyr Cys Arg Ala Asp Asn Gly His Val Pro Ile Gln Ser Lys Val Val Asn Ile Pro Val Arg Ile Pro Val Ser Arg Pro Val Leu Thr Leu Arg Ser Pro Gly A1a Gln Ala Ala Val Gly Asp Leu Leu G1u Leu His Cys Glu Ala Leu Arg Gly Ser Pro Pro Ile Leu Tyr Gln Phe Tyr His Glu Asp Val Thr Leu Gly Asn 5er Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Ala Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asn Asn Gly Leu Gly Ala Gln Cys Ser Glu Ala Val Pro Val Ser Ile Ser Gly Pro Asp Gly Tyr Arg Arg Asp Leu Met Thr Ala <210> 23 <211> 140 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 23 His Tyr Ala Arg Ala Arg Arg Lys Pro Gly Gly Leu Ser A1a Thr Gly Thr Ser Ser His Ser Pro Ser Glu Cys Gln Glu Pro 5er Ser Ser Arg Pro Ser Arg Ile Asp Pro Gln Glu Pro Thr His Ser Lys Pro Leu A1a Pro Met Glu Leu Glu Pro Met Tyr Ser Asn Ala Asn Pro Gly Asp Ser Asn Pro Ile Tyr Ser Gln Tle Trp Ser Ile Gln His Thr Lys Glu Asn Ser Ala Asn Cys Pro Met Met His Gln Glu His Glu Glu Leu Thr Val Leu Tyr 5er Glu Leu Lys Lys Thr His Pro Asp Asp Ser Ala Gly Glu Ala Ser Ser Arg Gly Arg Ala His Glu G1u Asp Asp Glu Glu Asn Tyr Glu Asn Val Pro Arg Val Leu Leu Ala Ser Asp His <210> 24 <211> 554 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:lnote =
synthetic construct <400> 24 Gly Val Ala Pro Lys Ala Val Leu Leu Leu Asn Pro Pro Trp Ser Thr Ala Phe Lys Gly Glu Lys Val Ala Leu Ile Cys Ser Ser Ile Ser His Ser Leu Ala Gln Gly Asp Thr Tyr Trp Tyr His Asp Glu Lys Leu Leu Lys Ile Lys His Asp Lys Ile Gln Ile Thr Glu Pro Gly Asn Tyr G1n Cys Lys Thr Arg Gly Ser Ser Leu Ser Asp Ala Val His Val Glu Phe Ser Pro Asp Trp Leu Ile Leu Gln Ala Leu His Pro Val Phe Glu Gly Asp Asn Val Tle Leu Arg Cys Gln Gly Lys Asp Asn Lys Asn Thr His Gln Lys Val Tyr Tyr Lys Asp Gly Lys Gln Leu Pro Asn Ser Tyr Asn Leu Glu Lys Ile Thr Val Asn Ser Val Ser Arg Asp Asn Ser Lys Tyr His Cys Thr Ala Tyr Arg Lys Phe Tyr Ile Leu Asp Tle Glu Val Thr 5er Lys Pro Leu Asn Ile Gln Val Gln Glu Leu Phe Leu His Pro Val Leu Arg Ala Ser Ser Ser Thr Pro Ile Glu Gly Ser Pro Met Thr Leu Thr Cys Glu Thr Gln Leu Ser Pro Gln Arg Pro Asp Val Gln Leu Gln Phe Ser Leu Phe Arg Asp Ser G1n Thr Leu Gly Leu Gly Trp Ser Arg 5er Pro Arg Leu Gln Ile Pro Ala Met Trp Thr Glu Asp Ser G1y Ser Tyr Trp Cys Glu Val Glu Thr Val Thr His Ser Tle Lys Lys Arg Ser Leu Arg Ser Gln Ile Arg Va1 Gln Arg Val Pro Val Ser Asn Val Asn Leu G1u Ile Arg Pro Thr Gly Gly Gln Leu Ile Glu Gly Glu Asn Met Val Leu Ile Cys Ser Val Ala G1n Gly Ser Gly Thr Val Thr Phe Ser Trp His Lys Glu Gly Arg Val Arg Ser Leu Gly Arg Lys Thr Gln Arg _ Ser Leu Leu Ala Glu Leu His Val Leu Thr Val Lys Glu Ser Asp Ala Gly Arg Tyr Tyr Cys Ala A1a Asp Asn Val His Ser Pro Ile Leu Ser Thr Trp Tle Arg Val Thr Val Arg Ile Pro Val Ser His Pro Val Leu Thr Phe Arg Ala Pro Arg Ala His Thr Val Val Gly Asp Leu Leu Glu Leu His Cys Glu Ser Leu Arg Gly Ser Pro Pro Ile Leu Tyr Arg Phe Tyr His Glu Asp Val Thr Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly A1a Ser Phe Asn Leu Ser Leu Thr Ala Glu His Ser Gly Asn Tyr Ser Cys Asp Ala Asp Asn Gly Leu Gly Ala Gln His Ser His Gly Val Ser Leu Arg Val Thr Val Pro Val Ser Arg Pro Val Leu Thr Leu Arg Ala Pro Gly Ala Gln Ala Val Val Gly Asp Leu Leu Glu Leu His Cys Glu Ser Leu Arg Gly 5er Phe Pro Ile Leu Tyr Trp Phe Tyr His Glu Asp Asp Thr Leu Gly Asn Ile Ser Ala His Ser Gly Gly Gly A1a Ser Phe Asn Leu Ser Leu Thr Thr Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asp Asn Gly Leu Gly Ala Gln His Ser Lys Val Val Thr Leu Asn Val Thr Gly Thr Ser Arg Asn Arg Thr Gly <210> 25 <211> 717 , <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 25 Gly Va1 Ala Pro Lys Ala Val Leu Leu Leu Asn Pro Pro Trp Ser Thr Ala Phe Lys Gly Glu Lys Va1 Ala Leu Ile Cys Ser Ser Ile Ser His Ser Leu Ala Gln Gly Asp Thr Tyr Trp Tyr His Asp Glu Lys Leu Leu Lys Ile Lys His Asp Lys Ile Gln Ile Thr Glu Pro Gly Asn Tyr Gln Cys Lys Thr Arg Gly Ser Ser Leu Ser Asp Ala Val His Val Glu Phe Ser Pro Asp Trp Leu Ile Leu Gln Ala Leu His Pro Val Phe Glu Gly Asp Asn Val Ile Leu Arg Cys Gln Gly Lys Asp Asn Lys Asn Thr His Gln Lys Val Tyr Tyr Lys Asp G1y Lys Gln Leu Pro Asn Ser Tyr Asn Leu Glu Lys Ile Thr Val Asn Ser Val Ser Arg Asp Asn Ser Lys Tyr His Cys Thr Ala Tyr Arg Lys Phe Tyr Ile Leu Asp Ile Glu Val Thr Ser Lys Pro Leu Asn Ile Gln Val Gln Glu Leu Phe Leu His Pro Val Leu Arg Ala Ser Ser Ser Thr Pro Ile Glu Gly Ser Pro Met Thr Leu Thr Cys Glu Thr Gln Leu Ser Pro Gln Arg Pro Asp Val Gln Leu Gln Phe Ser Leu Phe Arg Asp Ser Gln Thr Leu Gly Leu Gly Trp Ser Arg Ser Pro Arg Leu Gln Ile Pro Ala Met Trp Thr Glu Asp 5er Gly 5er Tyr Trp Cys Glu Val Glu Thr Val Thr His Ser Ile Lys Lys Arg Ser Leu Arg Ser Gln Ile Arg Val Gln Arg Val Pro Val Ser Asn Val Asn Leu Glu Ile Arg Pro Thr Gly Gly Gln Leu Ile Glu Gly Glu Asn Met Val Leu Tle Cys Ser Val Ala Gln Gly Ser Gly Thr Val Thr Phe Ser Trp His Lys Glu Gly Arg Val Arg Ser Leu Gly Arg Lys Thr Gln Arg Ser Leu Leu Ala Glu Leu His Val Leu Thr Val Lys Glu Ser Asp Ala Gly Arg Tyr Tyr Cys Ala Ala Asp Asn Val His Ser Pro Ile Leu Ser Thr Trp Ile Arg Val Thr Val Arg Ile Pro Val Ser His Pro Val Leu Thr Phe Arg Ala Pro Arg Ala His Thr Val Val Gly Asp Leu Leu Glu Leu His Cys Glu Ser Leu Arg Gly Ser Pro Pro Ile Leu Tyr Arg Phe Tyr His Glu Asp Val Thr Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Ala Glu His Ser Gly Asn Tyr Ser Cys Asp Ala Asp Asn Gly Leu Gly Ala Gln His Ser His Gly Val Ser Leu Arg Val Thr Val Pro Val Ser Arg Pro Val Leu Thr Leu Arg Ala Pro Gly Ala Gln Ala Val Val Gly Asp Leu Leu G1u Leu His Cys Glu 5er Leu Arg Gly Ser Phe Pro I1e Leu Tyr Trp Phe Tyr His G1u Asp Asp Thr Leu Gly Asn Ile Ser Ala His Ser Gly Gly Gly Ala Ser Phe Asn Leu Ser Leu Thr Thr Glu His Ser Gly Asn Tyr Ser Cys Glu Ala Asp Asn Gly Leu Gly Ala Gln His Ser Lys Val Val Thr Leu Asn Va1 Thr Gly Thr Ser Arg Asn Arg Thr Gly Leu Thr Ala Ala Gly Ile Thr Gly Leu Va1 Leu Ser Ile Leu Val Leu A1a Ala Ala Ala Ala Leu Leu His Tyr Ala Arg Ala Arg Arg Lys Pro Gly Gly Leu Ser Ala Thr Gly Thr Ser Ser His Ser Pro Ser Glu Cys Gln Glu Pro Ser Ser Ser Arg Pro 5er Arg Ile Asp Pro Gln Glu Pro Thr His Ser Lys Pro Leu Ala Pro Met Glu Leu G1u Pro Met Tyr Ser Asn Ala Asn Pro Gly Asp Ser Asn Pro Ile Tyr Ser Gln Ile Trp Ser Ile Gln His Thr Lys Glu Asn Ser Ala Asn Cys Pro Met Met His G1n Glu His Glu G1u Leu Thr Val Leu Tyr Ser Glu Leu Lys Lys Thr His Pro Asp Asp Ser Ala Gly Glu Ala Ser Ser Arg Gly Arg Ala His Glu Glu Asp Asp Glu Glu Asn Tyr Glu Asn Val Pro Arg Val Leu Leu Ala Ser Asp His <210> 26 <211> 104 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 26 Arg Ser Trp Arg Lys Ala Gly Pro Leu Pro Ser Gln Ile Pro Pro Thr Ala Pro Gly Gly Glu Gln Cys Pro Leu Tyr Ala Asn Val His His Gln Lys Gly Lys Asp Glu Gly Val Val Tyr Ser Val Val His Arg Thr Ser Lys Arg Ser Glu Ala Arg Ser Ala Glu Phe Thr Val Gly Arg Lys Asp Ser Ser Ile Ile Cys Ala Glu Val Arg Cys Leu Gln Pro Ser Glu Val Ser Ser Thr G1u Val Asn Met Arg Ser Arg Thr Leu Gln Glu Pro Leu Ser Asp Cys Glu Glu Val Leu Cys <210> 27 <211> 291 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 27 Lys Thr Val Trp Leu Tyr Leu Gln Ala Trp Pro Asn Pro Val Phe Glu 1 5 10 l5 Gly Asp Ala Leu Thr Leu Arg Cys Gln Gly Trp Lys Asn Thr Pro Leu Ser Gln Val Lys Phe Tyr Arg Asp Gly Lys Phe Leu His Phe Ser Lys Glu Asn G1n Thr Leu Ser Met G1y Ala Ala Thr Val G1n Ser Arg Gly G1n Tyr Ser Cys Ser Gly Gln Val Met Tyr Ile Pro Gln Thr Phe Thr Gln Thr Ser Glu Thr Ala Met Val Gln Val Gln Glu Leu Phe Pro Pro Pro Val Leu 5er Ala Ile Pro Ser Pro Glu Pro Arg Glu Gly Ser Leu Val Thr Leu Arg Cys Gln Thr Lys Leu His Pro Leu Arg Ser Ala Leu Arg Leu Leu Phe Ser Phe His Lys Asp G1y His Thr Leu Gln Asp Arg Gly Pro His Pro Glu Leu Cys Ile Pro Gly A1a Lys Glu Gly Asp Ser Gly Leu Tyr Trp Cys Glu Va1 Ala Pro Glu Gly Gly Gln Val G1n Lys Gln Ser Pro Gln Leu Glu Val Arg Val Gln Ala Pro Val Ser Arg Pro Val Leu Thr Leu His His Gly Pro Ala Asp Pro Ala Val G1y Asp Met Va1 Gln Leu Leu Cys Glu Ala Gln Arg Gly Ser Pro Pro Ile Leu Tyr Ser Phe Tyr Leu Asp Glu Lys Ile Val Gly Asn His Ser Ala Pro Cys Gly Gly Thr Thr Ser Leu Leu Phe Pro Val Lys Ser Glu Gln Asp Ala Gly Asn Tyr Ser Cys Glu Ala Glu Asn Ser Val Ser Arg Glu Arg Ser Glu Pro Lys Lys Leu Ser Leu Lys Gly Ser Gln Val Leu Phe Thr Pro Ala Ser Asn <210> 28 <211> 919 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 28 Lys Thr Val Trp Leu Tyr Leu Gln Ala Trp Pro Asn Pro Val Phe Glu Gly Asp Ala Leu Thr Leu Arg Cys Gln Gly Trp Lys Asn Thr Pro Leu Ser Gln Val Lys Phe Tyr Arg Asp Gly Lys Phe Leu His Phe Ser Lys Glu Asn Gln Thr Leu Ser Met Gly Ala Ala Thr Val Gln Ser Arg Gly Gln Tyr Ser Cys Ser G1y Gln Val Met Tyr Ile Pro Gln Thr Phe Thr Gln Thr Ser Glu Thr Ala Met Val Gln Val Gln Glu Leu Phe Pro Pro Pro Val Leu Ser Ala Tle Pro Ser Pro Glu Pro Arg Glu Gly Ser Leu Val Thr Leu Arg Cys Gln Thr Lys Leu His Pro Leu Arg Ser Ala Leu Arg Leu Leu Phe Ser Phe His Lys Asp Gly His Thr Leu Gln Asp Arg Gly Pro His Pro Glu Leu Cys Ile Pro Gly Ala Lys Glu Gly Asp Ser Gly Leu Tyr Trp Cys Glu Val Ala Pro Glu Gly Gly Gln Val Gln Lys Gln Ser Pro Gln Leu Glu Va1 Arg Val Gln Ala Pro Val Ser Arg Pro Val Leu Thr Leu His His Gly Pro Ala Asp Pro Ala Val Gly Asp Met Val Gln Leu Leu Cys Glu Ala G1n Arg Gly Ser Pro Pro Ile Leu Tyr Ser Phe Tyr Leu Asp Glu Lys Tle Val Gly Asn His Ser Ala Pro Cys G1y Gly Thr Thr Ser Leu Leu Phe Pro Val Lys Ser Glu Gln Asp Ala Gly Asn Tyr Ser Cys Glu Ala Glu Asn Ser Val Ser Arg Glu Arg Ser Glu Pro Lys Lys Leu Ser Leu Lys Gly Ser Gln Val Leu Phe Thr Pro Ala Ser Asn Trp Leu Val Pro Trp Leu Pro Ala Ser Leu Leu Gly Leu Met Val Tle Ala Ala Ala Leu Leu Val Tyr Val Arg Ser Trp Arg Lys Ala Gly Pro Leu Pro Ser Gln Ile Pro Pro Thr Ala Pro Gly Gly Glu Gln Cys Pro Leu Tyr Ala Asn Val His His Gln Lys Gly Lys Asp Glu Gly Val Val Tyr Ser Val Val His Arg Thr Ser Lys Arg Ser Glu Ala Arg Ser Ala Glu Phe Thr Val Gly Arg Lys Asp Ser Ser Ile Tle Cys Ala Glu Val Arg Cys Leu Gln Pro Ser Glu Val 5er Ser Thr Glu Val Asn Met Arg Ser Arg Thr Leu Gln Glu Pro Leu Ser Asp Cys Glu Glu Val Leu Cys <210> 29 <211> 16 <212> PRT

<213> Artificial Sequence <220>

<223> Description of ArtificialSequence:/note =

synthetic construct <400> 29 Met Leu Pro Arg Leu Leu Ile Cys Ala Cys Glu Leu Leu Pro Leu Pro <210> 30 <211> 19 <212> PRT

<213> Artificial Sequence <220>

<223> Description of ArtificialSequence:/note =

synthetic construct <400> 30 Ser Leu Leu Val Ile Phe Asp Thr Glu Leu Tr Ala Val Gln t L

p 10 15 eu Me Ala Asp Ser <210> 31 <211> 17 <212> PRT
<2l3> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 31 Met Leu Leu Trp Leu Leu Leu Leu Ile Leu Thr Pro Gly Arg Glu Gln Ser <210> 32 <211> 15 <212> PRT

<213> Artificial Sequence <220>
<223> Description of Artificial 5equence:/note =
synthetic construct <400> 32 Met Leu Zeu Trp Thr Ala Val Zeu Zeu Phe Val Pro Cys Val Gly <210> 33 <211> 51 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 33 atgctgccga ggctgttgct gttgatctgt gctccactct gtgaacctgc c 51 <210> 34 <211> 1236 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 34 gagctgtttt tgatagccag cccctcccat cccacagagg ggagcccagt gaccctgacg 60 tgtaagatgc cctttctaca gagttcagat gcccagttcc agttctgctt tttcagagac 120 acccgggcct tgggcccagg ctggagcagc tcccccaagc tccagatcgc tgccatgtgg 180 aaagaagaca cagggtcata ctggtgcgag gcacagacaa tggcgtccaa agtcttgagg 240 agcaggagat cccagataaa tgtgcacagg gtccctgtcg ctgatgtgag cttggagact 300 cagcccccag gaggacaggt gatggaggga gacaggctgg tcctcatctg ctcagttgct 360 atgggcacag gagacatcac cttcctttgg tacaaagggg ctgtaggttt aaaccttcag 420 tcaaagaccc agcgttcact gacagcagag tatgagattc cttcagtgag ggagagtgat 480 gctgagcaat attactgtgt agctgaaaat ggctatggtc ccagccccag tgggctggtg 540 agcatcactg tcagaatccc ggtgtctcgc ccaatcctca tgctcagggc tcccagggcc 600 caggctgcag tggaggatgt gctggagctt cactgtgagg ccctgagagg ctctcctcca 660 atcctgtact ggttttatca cgaggatatc accctgggga gcaggtcggc cccctctgga 720 ggaggagcct ccttcaacct ttccctgact gaagaacatt ctggaaacta ctcctgtgag 780 gccaacaatg gcctgggggc ccagcgcagt gaggcggtga cactcaactt cacagtgcct 840 actggggcca gaagcaatca tcttacctca ggagtcattg aggggctgct cagcaccctt 900 ggtccagcca ccgtggcctt attattttgc tacggcctca aaagaaaaat aggaagacgt 960 tcagccaggg atccactcag gagccttccc agccctctac cccaagagtt cacctacctc 1020 aactcaccta ccccagggca gctacagcct atatatgaaa atgtgaatgt tgtaagtggg 1080 gatgaggttt attcactggc gtactataac cagccggagc aggaatcagt agcagcagaa 1140 accctgggga cacatatgga ggacaaggtt tccttagaca tctattccag gctgaggaaa 1200 gcaaacatta cagatgtgga ctatgaagat gctatg 1236 <210> 35 <211> 60 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =

synthetic construct <400>
35 ggtcatctttgatgcagtcactgaacaggcagattcgctg60 atgctgctgt ggtcattgct <210>

<211>

<212>
DNA

<213> icial nce Artif Seque <220>

<223> iption te =
Descr of Artificial Sequence:/no synth etic construct <400>
36 cgccctcttctgtcttcgaaggagacagcatcgttctgaaatgccaggga60 acccttgtgg gaacagaactggaaaattcagaagatggcttaccataaggataacaaagagttatctgtt120 ttcaaaaaattctcagatttccttatccaaagtgcagttttaagtgacagtggtaactat180 ttctgtagtaccaaaggacaactctttctctgggataaaacttcaaatatagtaaagata240 aaagtccaagagctctttcaacgtcctgtgctgactgccagctccttccagcccatcgaa300 gggggtccagtgagcctgaaatgtgagacccggctctctccacagaggttggatgttcaa360 ctccagttctgcttcttcagagaaaaccaggtcctggggtcaggctggagcagctctccg420 gagctccagatttctgccgtgtggagtgaagacacagggtcttactggtgcaaggcagaa480 acggtgactcacaggatcagaaaacagagcctccaatcccagattcacgtgcagagaatc540 cccatctctaatgtaagcttggagatccgggcccccgggggacaggtgactgaaggacaa600 aaactgatcctgctctgctcagtggctgggggtacaggaaatgtcacattctcctggtac660 agagaggccacaggaaccagtatgggaaagaaaacccagcgttccctgtcagcagagctg720 gagatcccagctgtgaaagagagtgatgccggcaaatattactgtagagctgacaacggc780 catgtgcctatccagagcaaggtggtgaatatccctgtgagaattccagtgtctcgccct840 gtcctcaccctcaggtctcctggggcccaggctgcagtgggggacctgctggagcttcac900 tgtgaggccctgagaggctctcccccaatcttgtaccaattttatcatgaggatgtcacc960 cttgggaacagctcggccccctctggaggaggggcctccttcaacctctctttgactgca1020 gaacattctggaaactactcctgtgaggccaacaacggcctgggggcccagtgcagtgag1080 gcagtgccagtctccatctcaggacctgatggctatagaagagacctcatgacagctgga1140 gttctctggggactgtttggtgtccttggtttcactggtgttgctttgctgttgtatgcc1200 ttgttccacaagatatcaggagaaagttctgccactaatgaacccagaggggcttccagg1260 ccaaatcctcaagagttcacctattcaagcccaaccccagacatggaggagctgcagcca1320 gtgtatgtcaatgtgggctctgtagatgtggatgtggtttattctcaggtctggagcatg1380 cagcagccagaaagctcagcaaacatcaggacacttctggagaacaaggactcccaagtc1440 atctactcttctgtgaagaaatca 1464 <210>

<211>

<212>
DNA

<213> ficial Arti Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 37 atgcttctgt ggctgctgct gctgatcctg actcctggaa gagaacaatc aggg 54 <210> 38 <211> 2148 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
38 tctcctcaatcctccatggtccacagccttcaaaggagaa60 gtggccccaa aagctgtact aaagtggctctcatatgcagcagcatatcacattccctagcccagggagacacatattgg120 tatcacgatgagaagttgttgaaaataaaacatgacaagatccaaattacagagcctgga180 aattaccaatgtaagacccgaggatcctccctcagtgatgccgtgcatgtggaattttca240 cccgactggctgatcctgcaggctttacatcctgtctttgaaggagacaatgtcattctg300 agatgtcaggggaaagacaacaaaaacactcatcaaaaggtttactacaaggatggaaaa360 cagcttcctaatagttataatttagagaagatcacagtgaattcagtctccagggataat420 agcaaatatcattgtactgcttataggaagttttacatacttgacattgaagtaacttca480 aaacccctaaatatccaagttcaagagctgtttctacatcctgtgctgagagccagctct540 tccacgcccatagaggggagtcccatgaccctgacctgtgagacccagctctctccacag600 aggccagatgtccagctgcaattctccctcttcagagatagccagaccctcggattgggc660 tggagcaggtcccccagactccagatccctgccatgtggactgaagactcagggtcttac720 tggtgtgaggtggagacagtgactcacagcatcaaaaaaaggagcctgagatctcagata780 cgtgtacagagagtccctgtgtctaatgtgaatctagagatccggcccaccggagggcag840 ctgattgaaggagaaaatatggtccttatttgctcagtagcccagggttcagggactgtc900 acattctcctggcacaaagaaggaagagtaagaagcctgggtagaaagacccagcgttcc960 ctgttggcagagctgcatgttctcaccgtgaaggagagtgatgcagggagatactactgt1020 gcagctgataacgttcacagccccatcctcagcacgtggattcgagtcaccgtgagaatt1080 ccggtatctcaccctgtcctcaccttcagggctcccagggcccacactgtggtgggggac1140 ctgctggagcttcactgtgagtccctgagaggctctcccccgatcctgtaccgattttat1200 catgaggacgtcaccctggggaacagctcagccccctctggaggaggagcctccttcaac1260 ctctctctgactgcagaacattctggaaactactcctgtgatgcagacaatggcctgggg1320 gcccagcacagtcatggagtgagtctcagggtcacagttccggtgtctcgccccgtcctc1380 accctcagggctcccggggcccaggctgtggtgggggacctgctggagcttcactgtgag1440 tccctgagaggctccttcccgatcctgtactggttttatcacgaggatgacaccttgggg1500 aacatctcggcccactctggaggaggggcatccttcaacctctctctgactacagaacat1560 tctggaaactactcatgtgaggctgacaatggcctgggggcccagcacagtaaagtggtg1620 acactcaatgttacaggaacttccaggaacagaacaggccttaccgctgcgggaatcacg1680 gggctggtgctcagcatcctcgtccttgctgctgctgctgctctgctgcattacgccagg1740 gcccgaaggaaaccaggaggactttctgccactggaacatctagtcacagtcctagcgag1800 tgtcaggagccttcctcgtccaggccttccaggatagaccctcaagagcccactcactct1860 aaaccactagccccaatggagctggagccaatgtacagcaatgcaaatcctggagatagc1920 aacccgatttattcccagatctggagcatccagcatacaaaagaaaactcagctaattgt1980 ccaatgatgcatcaagagcatgaggaacttacagtcctctattcagaactgaagaagaca2040 cacccagacgactctgcaggggaggctagcagcagaggcagggcccatgaagaagatgat2100 gaagaaaactatgagaatgtaccacgtgtattactggcctcagaccac 2148 <210> 39 <211> 48 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 39 atgctgctct ggacggctgt gctgctcttt gttccctgtg ttgggaaa 48 <210> 40 <211> 2003 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 40 aggtctggttgctctctgccggcttcgccctgacctgtttctgacctgtgttccctccgc60 tgtgccagaacaggccccatgctgctctggacggctgtgctgctctttgttccctgtgtt120 gggaaaactgtctggctgtacctccaagcctggccaaaccctgtgtttgaaggagatgcc180 ctgactctgcgatgtcagggatggaagaatacaccactgtctcaggtgaagttctacaga240 gatggaaaattccttcatttctctaaggaaaaccagactctgtccatgggagcagcaaca300 gtgcagagccgtggccagtacagctgctctgggcaggtgatgtatattccacagacattc360 acacaaacttcagagactgccatggttcaagtccaagagctgtttccacctcctgtgctg420 agtgccatcccctctcctgagccccgagagggtagcctggtgaccctgagatgtcagaca480 aagctgcaccccctgaggtcagccttgaggctccttttctccttccacaaggacggccac540 accttgcaggacaggggccctcacccagaactctgcatcccgggagccaaggagggagac600 tctgggctttactggtgtgaggtggcccctgagggtggccaggtccagaagcagagcccc660 cagctggaggtcagagtgcaggctcctgtatcccgtcctgtgctcactctgcaccacggg720 cctgctgaccctgctgtgggggacatggtgcagctcctctgtgaggcacagaggggctcc780 cctccgatcctgtattccttctaccttgatgagaagattgtggggaaccactcagctccc840 tgtggtggaaccacctccctcctcttcccagtgaagtcagaacaggatgctgggaactac900 tcctgcgaggctgagaacagtgtctccagagagaggagtgagcccaagaagctgtctctg960 aagggttctcaagtcttgttcactcccgccagcaactggctggttccttggcttcctgcg1020 agcctgcttggcctgatggttattgctgctgcacttctggtttatgtgagatcctggaga1080 aaagctgggccccttccatcccagataccacccacagctccaggtggagagcagtgccca1140 ctatatgccaacgtgcatcaccagaaagggaaagatgaaggtgttgtctactctgtggtg1200 catagaacctcaaagaggagtgaagccaggtctgctgagttcaccgtggggagaaaggac1260 agttctatcatctgtgcggaggtgagatgcctgcagcccagtgaggtttcatccacggag1320 gtgaatatgagaagcaggactctccaagaaccccttagcgactgtgaggaggttctctgc1380 tagtgatggtgttctcctatcaacacacgcccacccccagtctccagtgctcctcaggaa1440 gacagtggggtcctcaactctttctgtgggtccttcagttcccaagcccagcatcacaga1500 gccccctgagcccttgtcctggtcaggagcacctgaaccctgggttcttttcttagcaga1560 agaccaaccaatggaatgggaagggagatgctcccaccaacacacacacttaggttcaat1620 cagtgacactggacacataagccacagatgtcttctttccatacaagcatgttagttcgc1680 cccaatatacatatatatatgaaatagtcatgtgccgcataacaacatttcagtcagtga1740 tagactgcatacacaacagtggtcccataagactgtaatggagtttaaaaattcctactg1800 cctagtgatatcatagttgccttaacatcataacacaacacatttctcacgcgtttgtgg1860 tgatgctggtacaaacaagctacagcgccgctagtcatatacaaatatagcacatacaat1920 tatgtacagtacactatacttgataatgataataaacaactatgttactggtctaaaaaa1980 aaaaaaaaaaaaaaaaaaaaaaa <210> 41 <211> 23 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 41 tgagtctcag ggtcacagtt ccg 23 <210> 42 <211> 26 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 42 gctcttgaac ttggatattt aggggt 26 <210> 43 <211> 25 <212> DNA

<213> Artificial Sequence <220>

<223> Description of ArtificialSequence:/note =

synthetic construct <400> 43 ccagtgtatg tcaatgtggg ctctg <210> 44 <211> 27 r <212> DNA

<213> Artificial Sequence <220>

<223> Description of ArtificialSequence:/note =

synthetic construct <400> 44 cgttgaaaga gctcttggac ttttatc <210> 45 <211> 27 <212> DNA

<213> Artificial Sequence <220>

<223> Description of ArtificialSequence:/note =

synthetic construct <400> 45 27 gcctcaaaag aaaaatagga agacgtt <210> 46 <211> 23 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 46 aagctcacat cagcgacagg gac <210> 47 <211> 22 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 47 tcttggagat aagtcgggct tt <210> 48 <211> 25 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 48 atcctgcagc ccagcctcgt aggag <210> 49 <211> 24 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 49 ggtcctcatg ctgctgtggt catt <210> 50 <211> 24 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 50 ~ 24 gctgttgatc ttcccttctg attc <210> 51 <211> 24 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 51 atgctgccga ggctgttgct gttg <210> 52 <211> 24 <212> DNA

<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 52 catagcatct tcatagtcca catc <210> 53 <211> 24 <212> DNA

<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 53 ctcaacttca cagtgcctac tggg 24 <210> 54 <211> 24 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 54 tcctgcagag tcactaacct tgag 24 <210> 55 <211> 25 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct ' <400> 55 ccagtgtatg tcaatgtggg ctctg 25 <210> 56 <211> 24 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 56 cattcttccc tcaaatcttt acac 24 <210> 57 <211> 21 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 57 cagcacgtgg attcgagtca c 21 <210> 58 <211> 24 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 58 cagatctggg aataaatcgg gttg 24 <210> 59 <211> 21 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 59 tcttcagaga tggcgaggtc a 21 <210> 60 <211> 26 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 60 ttttggggtg tacatcaaca tacaag 26 <210> 61 <211> 24 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note synthetic construct <400> 61 tgttgccctg tttcttccaa taca 24 <210> 62 <2l1> 20 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 62 cagagttggc cgacctacgc 20 <210> 63 <211> 32 <212> PRT
<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =
synthetic construct <221> VARIANT
<222> 5, 15, 17, 22, 28 <223> X can be any amino acid <400> 63 Gly Glu Pro Ile Xaa Leu Arg Cys His Ser Trp Lys Asp Lys Xaa Leu Xaa Lys Val Thr Tyr Xaa Gln Asn Gly Lys Ala Xaa Lys Phe Phe His <210> 64 <211> 17 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <221> VARIANT
<222> 1 <223> X can be either Glu or Asp <221> VARIANT
<222> 7 <223> X can be either Leu or Ile <221> VARIANT
<222> 17 <223> X can be either Leu or Ile <221> VARIANT
<222> 2-3, 5-6, 8-13, 15-16 <223> X can be any amino acid <400> 64 Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Xaa <210> 65 - <211> 18 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <221> VARIANT
<222> 1 <223> X can be either Glu or Asp <221> VARIANT
<222> 7 <223> X can be either Leu or Ile <221> VARIANT
<222> 18 ' <223> X can be either Leu or Ile <221> VARIANT
<222> 2-3, 5-6, 8-14, 16-17 <223> X can be any amino acid <400> 65 Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Xaa <210> 66 <211> 19 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <221> VARIANT
<222> 1 <223> X can be either Glu or Asp <221> VARIANT
<222> 7 <223> X can be either Leu or Ile <221> VARIANT
<222> 19 <223> X can be either Leu or Ile <221> VARIANT
<222> 2-3, 5-6, 8-15, 17-18 <223> X can be any amino acid <400> 66 Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Xaa <210> 67 <211> 6 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <221> VARIANT
<222> 1 <223> X can be either Ile orVal or Leu or Ser <221> VARIANT
<222> 2, 4-5 <223> X can be any amino acid <221> VARIANT
<222> 6 <223> X can be Leu or Val <400> 67 Xaa Xaa Tyr Xaa Xaa Xaa <210> 68 <211> 492 <212> PRT
<213> Artificial Sequenoe <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 68 Asp Trp Leu Ser Tle Ser Leu Pro His Arg Ser Tyr Glu Gly Asp Gln 1 5 l0 15 Val Val Ile Ser Cys Thr Gly Lys Asn Asn Gly Asp Ile Lys Arg Leu Lys Tyr Phe Lys Asp Gly Tyr His Ile Glu Thr Tyr Ser Ser Ala Ser Ser Tyr Thr Ile Arg Asn Ala Arg Arg Gly Asp Ser Gly Ser Tyr Ser Cys Lys Ala Asp Arg Lys Phe Phe Leu Phe Ile Asp Thr Thr Glu Glu Thr Gly Ser Lys Trp Leu Asn Val G1n Glu Leu Phe Pro Ala Pro Gly Leu Thr A1a Ser Pro Leu Gln Pro Val Glu Gly Ser Ser Va1 Thr Leu Ser Cys Asn Thr Trp Leu Pro Ser Asp Arg Ala Thr Thr Gln Leu Arg Tyr Ser Phe Phe Lys Asp Gly His Thr Leu Gln Ser Gly Trp Thr Ser Ser Lys Phe Thr Ile Ser Ala Ile Ser Lys Glu Asp Ser Gly Asn Tyr Trp Cys Glu Ala Met Thr Ala Ser Arg Ser Val Ser Lys Gln Ser His Arg Ser Tyr Ile Asp Val Glu Arg Ile Pro Val Ser Gln Va1 Thr Met 6 Glu Ile Gln Pro Ser Arg Gly Trp Gly Val Glu Gly Glu Pro Leu Val Val Glu Gly G1u Pro Leu Val Leu Ala Cys Ser Val Ala Lys Gly Thr Gly Leu Ile Thr Phe Ser Trp His Arg Gln Asp Thr Lys Glu Ser Va1 Gly Lys Lys Ser Gln Arg Ser Gln Arg Val Glu Leu G1u Ile Pro Thr Ile Arg Glu Ser His Ala Gly Gly Tyr Tyr Cys Thr Ala Asp Asn Asn Tyr Gly Leu Ile Gln Ser Ala Ile Va1 Asn Ile Thr Val Lys Ile Pro Val Leu Asn Pro Leu Leu Ser Ile Ser Val Pro Gly Val Leu Pro Phe Ile Gly Asp Val Ala Glu Leu His Cys Glu Asp Lys Arg A1a Ser Pro Pro Val Leu Tyr Trp Phe Tyr His Glu Asn Ile Thr Leu Ala Asn Thr Ser Ala Pro Phe Gly Gly Lys A1a Ser Phe Lys Leu Ser Leu Thr A1a Gly His Ser Gly Asn Tyr Ser Cys Glu Ala Glu Asn Ala Trp Gly Thr Lys Arg Ser Glu Val Val Thr Leu Asn Val Thr Glu Pro Pro Pro Lys Val Arg Leu Val Asn Gly Pro His His Cys Glu Gly Arg Val Glu Val Glu Gln Glu Gly Arg Trp Gly Thr Val Cys Asp Asp Gly Trp Asp Met Arg Asp Val Ala Val Val Cys Arg Glu Leu Gly Cys Gly Ala Ala Gln His Thr Pro Ile Ala Met Leu Tyr Pro Pro Ala Val Asp Glu Ala Leu Pro Val Leu Ile Gln Val Ala Leu Cys Asn Gly Thr Glu Lys Thr Leu Ala Glu Cys Asp Gln Val Glu Ala Phe Asp Cys Gly His Asp Glu Asp Ala Gly Ala Val Cys Glu Va1 Leu Pro Ser Thr Phe <210> 69 <211> 17 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 69 Met Pro Leu Cys Leu Leu Leu Leu Val Phe Ala Pro Val Gly Val Gln Ser <210> 70 <211> 383 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 70 Asp Trp Leu Ser Ile Ser Leu Pro His Arg Ser Tyr Glu Gly Asp Gln Val Val Ile Ser Cys Thr Gly Lys Asn Asn Gly Asp Tle Lys Arg Leu Lys Tyr Phe Lys Asp Gly Tyr His Ile Glu Thr Tyr Ser Ser Ala Ser Ser Tyr Thr Ile Arg Asn Ala Arg Arg Gly Asp Ser Gly Ser Tyr Ser Cys Lys Ala Asp Arg Lys Phe Phe Leu Phe Ile Asp Thr Thr Glu Glu Thr Gly Ser Lys Trp Leu Asn Va1 Gln Glu Leu Phe Pro Ala Pro Gly Leu Thr Ala 5er Pro Leu Gln Pro Val Glu Gly Ser Ser Val Thr Leu Ser Cys Asn Thr Trp Leu Pro Ser Asp Arg Ala Thr Thr Gln Leu Arg Tyr Ser Phe Phe Lys Asp Gly His Thr Leu Gln Ser Gly Trp Thr Ser 130 135 140 , Ser Lys Phe Thr Ile Ser Ala Ile 5er Lys Glu Asp Ser Gly Asn Tyr Trp Cys Glu Ala Met Thr Ala Ser Arg Ser Val Ser Lys Gln Ser His Arg Ser Tyr Ile Asp Val Glu Arg Ile Pro Val Ser Gln Val Thr Met Glu Ile Gln Pro Ser Arg Gly Trp Gly Val Glu Gly Glu Pro Leu Val Val Glu Gly Glu Pro Leu Val Leu Ala Cys Ser Val Ala Lys Gly Thr Gly Leu Ile Thr Phe Ser Trp His Arg Gln Asp Thr Lys Glu Ser Val Gly Lys Lys 5er Gln Arg Ser Gln Arg Val Glu Leu Glu Ile Pro Thr Tle Arg Glu Gly His Ala Gly Gly Tyr Tyr Cys Thr Ala Asp Asn Asn Tyr Gly Leu Tle Gln Ser Ala Ile Val Asn Ile Thr Val Lys Ile Pro _ 275 280 285 Val Leu Asn Pro Leu Leu Ser Ile Ser Val Pro Gly Val Leu Pro Phe Ile Gly Asp Val Ala Glu Leu His Cys Glu Asp Lys Arg Ala Ser Pro Pro Val Leu Tyr Trp Phe Tyr His Glu Asn Ile Thr Leu Ala Asn Thr Ser Ala Pro Phe Gly Gly Lys Ala Ser Phe Lys Leu Ser Leu Thr Ala Gly His Ser Gly Asn Tyr Ser Cys Glu Ala Glu Asn Ala Trp Gly Thr Lys Arg Ser Glu Val Val Thr Leu Asn Val Thr Gly Arg Thr Ile <210> 71 <211> 17 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 71 Met Pro Leu Cys Leu Leu Leu Leu Val Phe Ala Pro Val Gly Val Gln Ser <210> 72 <211> 17 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial 5equence:/note =
synthetic construct <400> 72 Met Leu Pro Trp Leu Leu Leu Leu Ile Cys Ala Leu Pro Cys Glu Pro Ala <210> 73 <211> 326 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 73 Gly Ile Ser Asp Val Ser Leu Lys Thr Arg Pro Pro Gly Gly Trp Val Met Glu Gly Asp Lys Leu Val Leu Ile Cys Ser Val Asp Arg Val Thr Gly Asn T1e Thr Tyr Phe Trp Tyr Arg Gly Ala Leu Gly Phe Gln Leu Glu Thr Lys Thr Gln Pro Ser Leu Thr Ala Glu Phe Glu Ile Ser Asp Met Lys Gln Ser Asp Ala Asp Gln Tyr Tyr Cys Ala A1a Asn Asp Gly His Asp Pro Ile Ala Ser Glu Leu Val Ser Ile His Val Arg Val Pro Val Ser Arg Pro Val Leu Thr Phe Gly Asp Ser Gly Thr Gln Ala Val Leu Gly Asp Leu Val Glu Leu His Cys Lys Ala Leu Arg Gly Ser Pro Pro Ile Phe Tyr Gln Phe Tyr His Glu Ser Ile I1e Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Phe Ser Leu Thr Ala Glu His Ser Gly Asn Phe Ser Cys Glu Ala Ser Asn Gly Gln Gly Ala Gln Arg Ser Glu Val Val A1a Leu Asn Leu Thr Gly Leu Ser Leu Val Pro Thr G1u Asn Gly Ile Ser His Leu Ser Leu Gly Leu Thr G1y Trp Leu Leu Gly Cys Leu Sex Pro Ile Thr Met Ala Leu Ile Phe Cys Tyr Trp Leu Lys Arg Lys Ile Gly Arg Gln Ser Glu Asp Pro Val Arg Ser Pro Pro Gln Thr Val Leu Gln Gly Ser Thr Tyr Pro Lys Ser Pro Asp Ser Arg G1n Pro Glu Pro Leu Tyr Glu Asn Va1 Asn Val Val Ser Gly Asn Glu Val Tyr Ser Leu Va1 Tyr His Thr Pro Gln Val Leu Glu Pro Ala Ala Ala Gln His Val Arg Thr His Gly Val Ser Glu Ser Phe Gln Val Ser Ser Gly Leu Tyr Ser Lys Pro Arg Ile Asn Ile Ala His Met Asp Tyr Glu Asp Ala Met <210> 74 <211> 203 <212> PRT

<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 74 Gly Ile Ser Asp Val Ser Leu Lys Thr Arg Pro Pro Gly Gly Trp Val Met Glu Gly Asp Lys Leu Val Leu Ile Cys Ser Val Asp Arg Val Thr Gly Asn Ile Thr Tyr'Phe Trp Tyr Arg Gly Ala Leu Gly Phe Gln Leu Glu Thr Lys Thr Gln Pro Ser Leu Thr Ala Glu Phe Glu Ile Ser Asp Met Lys Gln 5er Asp Ala Asp Gln Tyr Tyr Cys Ala Ala Asn Asp Gly His Asp Pro Ile Ala Ser Glu Leu Val Ser Ile His Val Arg Val Pro Val Ser Arg Pro Val Leu Thr Phe Gly Asp Ser Gly Thr Gln Ala Val Leu Gly Asp Leu Val Glu Leu His Cys Lys Ala Leu Arg Gly Ser Pro Pro Ile Phe Tyr Gln Phe Tyr His Glu Ser Ile Ile Leu Gly Asn Ser Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Phe Ser Leu Thr Ala Glu His 5er Gly Asn Phe Ser Cys Glu Ala Ser Asn Gly Gln Gly Ala Gln Arg Ser Glu Val Val Ala Leu Asn Leu Thr Gly Leu Ser Leu Val Pro Thr Glu Asn Gly Ile Ser His Leu Ser Leu <210> 75 <211> 17 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 75 Met Leu Pro Trp Leu Leu Leu Leu Ile Cys Ala Leu Pro Cys Glu Pro Ala <210> 76 <211> 100 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 76 Lys Arg Lys Ile Gly Arg Gln Ser Glu Asp Pro Val Arg Ser Pro Pro Gln Thr Val Leu G1n Gly Ser Thr Tyr Pro Lys Ser Pro Asp Ser Arg Gln Pro G1u Pro Leu Tyr Glu Asn Val Asn Val Val Ser Gly Asn Glu Val Tyr Ser Leu Val Tyr His Thr Pro Gln Val Leu Glu Pro Ala Ala Ala Gln His Val Arg Thr His Gly Val Ser Glu Ser Phe Gln Val Ser Ser Gly Leu Tyr Ser Lys Pro Arg Tle Asn Ile Ala His Met Asp Tyr Glu Asp Ala Met <210> 77 <211> 283 <2l2> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 77 Gly Ile Ser Asp Val Ser Leu Lys Thr Arg Pro Pro Gly Gly Trp Val Met Glu Gly Asp Lys Leu Val Leu Tle Cys Ser Val Asp Arg Val Thr Gly Asn Ile Thr Tyr Phe Trp Tyr Arg Gly Ala Leu Gly Phe G1n Leu Glu Thr Lys Thr Gln Pro Ser Leu Thr Ala Glu Phe Glu Ile Ser Asp Met Lys Gln Ser Asp Ala Asp Gln Tyr Tyr Cys Ala Ala Asn Asp Gly His Asp Pro Ile Ala Ser Glu Leu Val Ser Ile His Val Arg Val Pro Val 5er Arg Pro Val Leu Thr Phe Gly Asp Ser Gly Thr Gln A1a Va1 Leu Gly Asp Leu Val Glu Leu His Cys Lys A1a Leu Arg Gly Ser Pro Pro Ile Phe Tyr Gln Phe Tyr His Glu Ser Ile Ile Leu Gly Asn Ser l30 135 140 Ser Ala Pro Ser Gly Gly Gly Ala Ser Phe Asn Phe Ser Leu Thr Ala Glu His Ser Gly Asn Phe Ser Cys Glu Ala Ser Asn Gly Gln Gly Ala 165 ' 170 175 Gln Arg Ser Glu Val Val Ala Leu Asn Leu Thr Gly Arg Gln Ser Glu Asp Pro Val Arg Ser Pro Pro G1n Thr Val Leu Gln Gly Ser Thr Tyr Pro Lys Ser Pro Asp Ser Arg Gln Pro Glu Pro Leu Tyr Glu Asn Val Asn Val Val Ser Gly Asn Glu Va1 Tyr Ser Leu Va1 Tyr His Thr Pro Gln Val Leu Glu Pro A1a Ala Ala Gln His Val Arg Thr His Gly Val Ser Glu Ser Phe Gln Val Ser Ser Gly Leu Tyr Ser Lys Pro Arg Ile Asn Ile Ala His Met Asp Tyr Glu Asp Ala Met <210> 78 <211> 570 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 78 Gly Gln His Glu Ala Ala Gln Gln Ser Val Val Ser Leu Gln Pro Pro Trp Thr Thr Phe Phe Arg Gly Glu Val Val Thr Leu Thr Cys Tyr Arg Phe Gly Phe Ser Val Pro Gln Lys Thr Lys Trp Tyr Gln Lys Arg Lys Thr Val Lys Gln Thr Pro Gly Ala Leu Val Ile Lys Ala His Thr Leu Lys Val His Glu Ser Gly Glu Tyr Trp Cys Gln A1a Asp Ser Leu Leu Pro Ser Met His Val Asn Val Glu Phe Ser Glu Asp Phe Leu Val Leu Gln Ala Pro Pro Ala Val Phe Glu Gly Asp Ser Val Val Leu Arg Cys Tyr Ala Lys Lys Gly Ile Glu Ala Glu Thr Leu Thr Phe Tyr Lys Asp Gly Lys Ala Leu Thr Leu His His Gln Ser Glu Leu Ser Tle His His Ala Asn Leu Lys Asp Asn Gly Gln Tyr Lys Cys Thr Ser Lys Lys Lys Trp Ser Phe Gly Ser Leu Tyr Thr Ser Asn Thr Val Gly Val Gln Val Gln Glu Leu Phe Pro Arg Pro Val Leu Arg Ala Arg Pro Ser His Pro Ile Asp Gly Ser Pro Val Thr Leu Thr Cys Gln Thr Gln Leu Ser Ala Gln Lys Ser Asp A1a Arg Leu Gln Phe Cys Phe Phe Arg Asn Leu Gln Leu Leu Gly Ser Gly Cys Ser Arg Ser Ser Glu Phe His I1e Pro Ala Ile Trp Thr Glu Glu Ser Arg Arg Tyr Gln Cys Lys Ala Glu Thr Val Asn Ser Gln Val Arg Lys Gln Ser Thr Ala Phe Ile Ile Pro Val Gln Arg Ala Ser Ala Arg Phe G1n Thr His Ile Ile Pro Ala Ser Lys Leu Val Phe Glu Gly Gln Leu Leu Leu Leu Asn Cys Ser Val Lys Gly Val 2g0 295 300 Pro Gly Pro Leu Lys Phe 5er Trp Tyr Lys Lys Asp Met Leu Asn Glu Glu Thr Lys Ile Leu Lys Ser Ser Asn Ala Glu Phe Lys Ile Ser Gln 325 330 . 335 Val Asn Ile Ser Asp Ala Gly Glu Tyr His Cys Glu Ala Thr Asn Ser Arg Arg Ser Phe Val Ser Arg Ala Phe Pro Ile Thr Ile Lys Val Pro Val Ser G1n Pro Val Leu Thr Leu Ser Thr Gly Lys Thr G1n Ala Leu Glu Gly Asp Leu Met Thr Leu His Cys G1n 5er Gln Arg Gly Ser Pro Cys Ile Leu Tyr Glu Phe Phe Tyr Glu Asn Val Ser Leu G1y Asn Ser Ser Ile Leu Ser Gly Gly Gly Ala Tyr Phe Asn Phe Ser Met Ser Thr Glu Arg Ser Gly Asn Tyr Tyr Cys Thr Ala Asp Asn Gly Leu Gly Ala Gln Cys Ser Glu Ala Tle Arg Ile Ser Ile Phe Asp Met Thr Lys Asn Arg Ser Val Pro Met Ala Ala Gly Ile Thr Val Gly Leu Leu Ile Met Ala Val Gly Val Phe Leu Phe Tyr Cys Trp Phe Ser Arg Lys Ala Gly Gly Lys Pro Thr Ser Asp Asp Ser Arg Asn Pro Ser Asp Ser Glu Pro Gln Glu Pro Thr Tyr Tyr Asn Val Pro Ala Cys Ile Glu Leu Gln Pro Val Tyr Ser Asn Glu Pro Glu Glu Asn Val Ile Tyr Thr Glu Val Arg Arg Thr Gln Pro Arg Gln Lys His Ala Asp Gln Glu Ser Glu Ser Pro Arg Ser Arg Cys Gln Met Ala Glu Lys Lys <210> 79 <211> 25 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 79 Met Ser Gly Ser Phe Ser Pro Cys Val Val Phe Thr G1n Met Trp Leu Thr Leu Leu Val Val Thr Pro Val Asn <210> 80 <211> 468 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 80 Gly Gln His Glu Ala Ala Gln Gln Ser Val Val Ser Leu Gln Pro Pro Trp Thr Thr Phe Phe Arg Gly Glu Val Val Thr Leu Thr Cys Tyr Arg Phe Gly Phe Ser Val Pro Gln Lys Thr Lys Trp Tyr Gln Lys Arg Lys Thr Val Lys Gln Thr Pro Gly Ala Leu Val Ile Lys A1a His Thr Leu Lys Val His Glu Ser Gly Glu Tyr Trp Cys Gln Ala Asp Ser Leu Leu Pro Ser Met His Val Asn Val Glu Phe Ser Glu Asp Phe Leu Val Leu Gln Ala Pro Pro Ala Val Phe Glu Gly Asp Ser Val Val Leu Arg Cys Tyr Ala Lys Lys Gly Ile Glu Ala Glu Thr Leu Thr Phe Tyr Lys Asp Gly Lys Ala Leu Thr Leu His His Gln Ser Glu Leu Ser Ile His His Ala Asn Leu Lys Asp Asn Gly Gln Tyr Lys Cys Thr Ser Lys Lys Lys Trp Ser Phe Gly 5er Leu Tyr Thr Ser Asn Thr Va1 Gly Val Gln Val Gln Glu Leu Phe Pro Arg Pro Val Leu Arg Ala Arg Pro Ser His Pro Ile Asp Gly Ser Pro Val Thr Leu Thr Cys Gln Thr Gln Leu Ser Ala Gln Lys Ser Asp Ala Arg Leu Gln Phe Cys Phe Phe Arg Asn Leu Gln Leu Leu Gly Ser Gly Cys Ser Arg Ser Ser Glu Phe His Ile Pro Ala Ile Trp Thr Glu G1u 5er Arg Arg Tyr Gln Cys Lys Ala Glu Thr Val Asn Ser Gln Val Arg Lys Gln Ser Thr Ala Phe Ile Ile Pro Val Gln Arg Ala Ser Ala Arg Phe Gln Thr His Ile Ile Pro Ala 5er Lys Leu Val Phe Glu Gly Gln Leu Leu Leu Leu Asn Cys Ser Val Lys Gly Val Pro Gly Pro Leu Lys Phe Ser Trp Tyr Lys Lys Asp Met Leu Asn Glu Glu Thr Lys Ile Leu Lys Ser Ser Asn Ala Glu Phe Lys I1e Ser Gln Val Asn Ile Ser Asp Ala Gly Glu Tyr His Cys Glu Ala Thr Asn Ser Arg Arg Ser Phe Val Ser Arg Ala Phe Pro Ile Thr Ile Lys Val Pro Val Ser Gln Pro Val Leu Thr Leu Ser Thr Gly Lys Thr Gln Ala Leu Glu Gly Asp Leu Met Thr Leu His Cys Gln Ser Gln Arg Gly Ser Pro Cys Ile Leu Tyr G1u Phe Phe Tyr Glu Asn Val Ser Leu Gly Asn Ser Ser Ile Leu Ser Gly Gly Gly Ala Tyr Phe Asn Phe Ser Met Ser Thr Glu Arg Ser Gly Asn Tyr Tyr Cys Thr Ala Asp Asn Gly Leu Gly Ala Gln Cys Ser Glu Ala I1e Arg Ile Ser Ile Phe Asp Met Thr Lys Asn Arg 5er Val Pro <210> 81 <211> 79 <212> PRT
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 81 Ser Arg Lys Ala Gly Gly Lys Pro Thr Ser Asp Asp Ser Arg Asn Pro Ser Asp Ser Glu Pro Gln Glu Pro Thr Tyr Tyr Asn Val Pro Ala Cys Ile Glu Leu Gln Pro Val Tyr Ser Asn Glu Pro Glu Glu Asn Val Ile Tyr Thr G1u Val Arg Arg Thr Gln Pro Arg Gln Lys His Ala Asp Gln Glu Ser Glu 5er Pro Arg Ser Arg Cys Gln Met Ala Glu Lys Lys <210> 82 <211> 1973 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
82 ctatcccagatccgtggtccatctgcccta ctgcacctgt60 ccacagtgtt aggacttgag ctcaaagggagctacttgcctctagtctcatgcctctgtgcttgctgcttctggtcttcg120 ctcctgtcggagtccagtccgactggttgagcatcagccttccacaccgttcttatgaag180 gagaccaagtagttataagctgcacaggaaaaaataatggtgacataaagagactgaagt240 acttcaaggatggatatcacatagaaacttacagcagtgcttcaagctacaccattagga300 atgcaagacgtggtgacagtggctcctattcctgtaaggcagataggaaatttttcctat360 ttatagacacaacagaagaaacaggatctaagtggctgaatgtccaagagctgtttccag420 cacctgggctgacagccagccccctgcagcccgtagaggggagttcagtgaccctgtcct480 gcaacacctggctcccttcagatagggcaacgacccagctacgctattccttcttcaaag540 atggccacactttgcaatcgggctggacctcatcaaaatttaccatctcagcaatatcga600 aggaagactcaggaaattactggtgtgaagcaatgactgcctctcgcagtgtctcaaagc660 agagtcaccggtcctacatagatgtagagaggatccctgtatctcaagtcaccatggaaa720 tccagccttcaaggggctggggagttgaaggggagccactggtcgttgaaggggagcccc780 tggtcctggcttgttctgtggctaaaggcaccgggctaatcacgttctoctggcataggc840 aggacactaaggaaagtgtggggaagaaaagtcagcgttcccagagagtggagctggaga900 tccctactatcagggaaggccatgctggggggtactactgcacagcagacaacaactacg960 gcctgatccagagcgcaatcgtgaacatcaccgtgaaaattccagtgttgaacccgctcc1020 tctccatcagtgttcctggggtcttgcccttcatcggagatgtggcggagcttcactgtg1080 aagacaagagagcatctcctccggttctctactggttttatcatgaaaatatcactctgg1140 ctaacacctcggcaccttttggaggaaaggcatcctttaagctctctctgactgcagggc1200 attctgggaactactcttgtgaggctgaaaacgcctggggtaccaagcgcagtgaggtgg1260 taacgctcaatgtcacagagcccccacccaaagtgcgtttggtgaatggcccccaccact1320 gtgaaggacgcgtagaggtggagcaggaaggtcgctggggcactgtatgtgatgatggct1380 gggacatgagggatgtggctgtggtgtgccgagagctgggctgtggagcagcccaacaca1440 cacctatagccatgctgtatccaccagcagttgatgaagctctgcctgtgctcattcagg1500 tagccctgtgcaatggcacagaaaagaccctggctgaatgtgaccaggttgaggcctttg1560 attgtggacatgatgaggatgctggagctgtgtgtgaagtcttacccagcactttctgaa1620 gatctagagaccagagaccatcagacctcctactttctgcactgggcctcacagccctca1680 cggtctgcagctcccagtggacttccagacttcagctgtggcttatccttcaagaggact1740 cgaaactatattaatctgctctgagataatgttccaacagctccaaagaaagcccgagtc1800 ccttgtccccagaggccaagcttggaaaaattgttcccctgtccaggttccctgcctttc1860 tagttccttcttgctatctccttgggcagatgcagaggtggcacaagtaaggatcacata1920 catgtgcctgggcttccatctggtagaatgtggtctaacaaagcacatac 1973 aac <210> 83 <211> 1530 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 83 atgcctctgtgcttgctgcttctggtcttcgctcctgtcggagtccagtccgactggttg60 agcatcagccttccacaccgttcttatgaaggagaccaagtagttataagctgcacagga120 aaaaataatggtgacataaagagactgaagtacttcaaggatggatatcacatagaaact180 tacagcagtgcttcaagctacaccattaggaatgcaagacgtggtgacagtggctcctat240 tcctgtaaggcagataggaaatttttcctatttatagacacaacagaagaaacaggatct300 aagtggctgaatgtccaagagctgtttccagcacctgggctgacagccagccccctgcag360 cccgtagaggggagttcagtgaccctgtcctgcaacacctggctcccttcagatagggca420 acgacccagctacgctattccttcttcaaagatggccacactttgcaatcgggctggacc480 tcatcaaaatttaccatctcagcaatatcgaaggaagactcaggaaattactggtgtgaa540 gcaatgactgcctctcgcagtgtctcaaagcagagtcaccggtcctacatagatgtagag600 aggatccctgtatctcaagtcaccatggaaatccagccttcaaggggctggggagttgaa660 ggggagccactggtcgttgaaggggagcccctggtcctggcttgttctgtggctaaaggc720 accgggctaatcacgttctcctggcataggcaggacactaaggaaagtgtggggaagaaa780 agtcagcgttcccagagagtggagctggagatccctactatcagggaaggccatgctggg840 gggtactactgcacagcagacaacaactacggcctgatccagagcgcaatcgtgaacatc900 accgtgaaaattccagtgttgaacccgctcctctccatcagtgttcctggggtcttgccc960 ttcatcggagatgtggcggagcttcactgtgaagacaagagagcatctcctccggttctc1020 tactggttttatcatgaaaatatcactctggctaacacctcggcaccttttggaggaaag1080 gcatcctttaagctctctctgactgcagggcattctgggaactactcttgtgaggctgaa1140 aacgcctggggtaccaagcgcagtgaggtggtaacgctcaatgtcacagagcccccaccc1200 aaagtgcgtttggtgaatggcccccaccactgtgaaggacgcgtagaggtggagcaggaa1260 ggtcgctggggcactgtatgtgatgatggctgggacatgagggatgtggctgtggtgtgc1320 cgagagctgggctgtggagcagcccaacacacacctatagccatgctgtatccaccagca1380 gttgatgaagctctgcctgtgctcattcaggtagccctgtgcaatggcacagaaaagacc1440 ctggctgaatgtgaccaggttgaggcctttgattgtggacatgatgaggatgctggagct1500 gtgtgtgaagtcttacccagcactttctga 1530 <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>

<223> iption equence:/note =
Descr of Artificial S

synth etic construct <400>
84 ctatcccagatccgtggtccatctgccctaaggacttgagctgcacctgt60 ccacagtgtt ctcaaagggagctacttgcctctagtctcatgcctctgtgcttgctgcttctggtcttcg120 ctcctgtcggagtccagtccgactggttgagcatcagccttccacaccgttcttatgaag180 gagaccaagtagttataagctgcacaggaaaaaataatggtgacataaagagactgaagt240 acttcaaggatggatatcacatagaaacttacagcagtgcttcaagctacaccattagga300 atgcaagacgtggtgacagtggctcctattcctgtaaggcagataggaaatttttcctat360 ttatagacacaacagaagaaacaggatctaagtggctgaatgtccaagagctgtttccag420 cacctgggctgacagccagccccctgcagcccgtagaggggagttcagtgaccctgtcct480 gcaacacctggctcccttcagatagggcaacgacccagctacgctattccttcttcaaag540 atggccacactttgcaatcgggctggacctcatcaaaatttaccatctcagcaatatcga600 aggaagactcaggaaattactggtgtgaagcaatgactgcctctcgcagtgtctcaaagc660 agagtcaccggtcctacatagatgtagagaggatccctgtatctcaagtcaccatggaaa720 tccagccttcaaggggctggggagttgaaggggagccactggtcgttgaaggggagcccc780 tggtcctggcttgttctgtggctaaaggcaccgggctaatcacgttctcctggcataggc840 aggacactaaggaaagtgtggggaagaaaagtcagcgttcccagagagtggagctggaga900 tccctactatcagggaaggccatgctggggggtactactgcacagcagacaacaactacg960 gcctgatccagagcgcaatcgtgaacatcaccgtgaaaattccagtgttgaacccgctcc1020 tctccatcagtgttcctggggtcttgcccttcatcggagatgtggcggagcttcactgtg1080 aagacaagagagcatctcctccggttctctactggttttatcatgaaaatatcactctgg1140 ctaacacctcggcaccttttggaggaaaggcatcctttaagctctctctgactgcagggc1200 attctgggaactactcttgtgaggctgaaaacgcctggggtaccaagcgcagtgaggtgg1260 taacgctcaatgtcacaggtaggacaatttaatgatccattccagggtgcaacttgcctt1320 ctggccatgcccttcttctctcccttgcacctgtacctcttggtctttgaa 1371 <210> 85 <211> 1203 <212> DNA
<213> Artificial Sequence <220>
<223>
Description of Artificial 5equence:/note =

synth etic construct <400>
85 gcttgctgcttctggtcttcgctcctgtcggagtccagtccgactggttg60 atgcctctgt agcatcagccttccacaccgttcttatgaaggagaccaagtagttataagctgcacagga120 aaaaataatggtgacataaagagactgaagtacttcaaggatggatatcacatagaaact180 tacagcagtgcttcaagctacaccattaggaatgcaagacgtggtgacagtggctcctat240 tcctgtaaggcagataggaaatttttcctatttatagacacaacagaagaaacaggatct300 aagtggctgaatgtccaagagctgtttccagcacctgggctgacagccagccccctgcag360 cccgtagaggggagttcagtgaccctgtcctgcaacacctggctcccttcagatagggca420 acgacccagctacgctattccttcttcaaagatggccacactttgcaatcgggctggacc480 tcatcaaaatttaccatctcagcaatatcgaaggaagactcaggaaattactggtgtgaa540 gcaatgactgcctctcgcagtgtctcaaagcagagtcaccggtcctacatagatgtagag600 aggatccctgtatctcaagtcaccatggaaatccagccttcaaggggctggggagttgaa660 ggggagccactggtcgttgaaggggagcccctggtcctggcttgttctgtggctaaaggc720 accgggctaatcacgttctcctggcataggcaggacactaaggaaagtgtggggaagaaa780 agtcagcgttcccagagagtggagctggagatccctactatcagggaaggccatgctggg840 gggtactactgcacagcagacaacaactacggcctgatccagagcgcaatcgtgaacatc900 accgtgaaaattccagtgttgaacccgctcctctccatcagtgttcctggggtcttgccc960 ttcatcggagatgtggcggagcttcactgtgaagacaagagagcatctcctccggttctc1020 tactggttttatcatgaaaatatcactctggctaacacctcggcaccttttggaggaaag1080 gcatcctttaagctctctctgactgcagggcattctgggaactactcttgtgaggctgaa1140 aacgcctggggtaccaagcgoagtgaggtggtaacgctcaatgtcacaggtaggacaatt1200 taa <210> 86 <211> 1479 <212> DNA
<213> Artificial Sequence <220>

<223> Description rtificialequence:/note =
of A S

synth etic construct <400> 86 gactggttgagcatcagccttccacaccgttcttatgaaggagaccaagtagttataagc60 tgcacaggaaaaaataatggtgacataaagagactgaagtacttcaaggatggatatcac120 atagaaacttacagcagtgcttcaagctacaccattaggaatgcaagacgtggtgacagt180 ggctcctattcctgtaaggcagataggaaatttttcctatttatagacacaacagaagaa240 acaggatctaagtggctgaatgtccaagagctgtttccagcacctgggctgacagccagc300 cccctgcagcccgtagaggggagttcagtgaccctgtcctgcaacacctggctcccttca360 gatagggcaacgacccagctacgctattccttcttcaaagatggccacactttgcaatcg420 ggctggacctcatcaaaatttaccatctcagcaatatcgaaggaagactcaggaaattac480 tggtgtgaagcaatgactgcctctcgcagtgtctcaaagcagagtcaccggtcctacata540 gatgtagagaggatccctgtatctcaagtcaccatggaaatccagccttcaaggggctgg600 ggagttgaaggggagccactggtcgttgaaggggagcccctggtcctggcttgttctgtg660 gctaaaggcaccgggctaatcacgttctcctggcataggcaggacactaaggaaagtgtg720 gggaagaaaagtcagcgttcccagagagtggagctggagatccctactatcagggaaggc780 catgctggggggtactactgcacagcagacaacaactacggcctgatccagagcgcaatc840 gtgaacatcaccgtgaaaattccagtgttgaacccgctcctctccatcagtgttcctggg900 gtcttgcccttcatcggagatgtggcggagcttcactgtgaagacaagagagcatctcct960 ccggttctctactggttttatcatgaaaatatcactctggctaacacctcggcacctttt1020 ' ggaggaaaggcatcctttaagctctctctgactgcagggcattctgggaactactcttgt1080 gaggctgaaaacgcctggggtaccaagcgcagtgaggtggtaacgctcaatgtcacagag1140 cccccacccaaagtgcgtttggtgaatggcccccaccactgtgaaggacgcgtagaggtg1200 aag gtcgctggggcactgtatgtgatgatggctgggacatgagggatgtggct1260 ca a gg gagagctgggctgtggagcagcccaacacacacctatagccatgctgtat1320 g g gtggtgtgcc ccaccagcagttgatgaagctctgcctgtgctcattcaggtagccctgtgcaatggcaca1380 accc tggctgaatgtgaccaggttgaggcctttgattgtggacatgatgaggat1440 aaaa g tgtgtgaagtcttacccagcactttctga 1479 g gctggagctg <210>

<211>

<212>
DNA

<213>
Artificial Sequence <220>
<223>
Description of Artificial Sequence:/note =

synth etic construct <400>
87 gcatcagccttccacaccgttcttatgaaggagaccaagtagttataagc60 gactggttga tgcacaggaaaaaataatggtgacataaagagactgaagtacttcaaggatggatatcac120 atagaaacttacagcagtgcttcaagctacaccattaggaatgcaagacgtggtgacagt180 ggctcctattcctgtaaggcagataggaaatttttcctatttatagacacaacagaagaa240 acaggatctaagtggctgaatgtccaagagctgtttccagcacctgggctgacagccagc300 cccctgcagcccgtagaggggagttcagtgaccctgtcctgcaacacctggctcccttca360 gatagggcaacgacccagctacgctattccttcttcaaagatggccacactttgcaatcg420 ggctggacctcatcaaaatttaccatctcagcaatatcgaaggaagactcaggaaattac480 tggtgtgaagcaatgactgcctctcgcagtgtctcaaagcagagtcaccggtcctacata540 gatgtagagaggatccctgtatctcaagtcaccatggaaatccagccttcaaggggctgg600 ggagttgaaggggagccactggtcgttgaaggggagcccctggtcctggcttgttctgtg660 gctaaaggcaccgggctaatcacgttctcctggcataggcaggacactaaggaaagtgtg720 gggaagaaaagtcagcgttcccagagagtggagctggagatccctactatcagggaaggc780 catgctggggggtactactgcacagcagacaacaactacggcctgatccagagcgcaatc840 gtgaacatcaccgtgaaaattccagtgttgaacccgctcctctccatcagtgttcctggg900 gtcttgcccttcatcggagatgtggcggagcttcactgtgaagacaagagagcatctcct960 ccggttctctactggttttatcatgaaaatatcactctggctaacacctcggcacctttt1020 ggaggaaaggcatcctttaagctctctctgactgcagggcattctgggaactactcttgt1080 gaggctgaaaacgcctggggtaccaagcgcagtgaggtggtaacgctcaatgtcacaggt1140 aggacaatttas <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>

<223> iption equence:/note =
Descr of Artificial S

synth etic construct <400>
88 ggttttgcttttttttcttttggtgagaggtaccttcaagttaccatccc60 tgagtagtct cagcctggtcctcatgctaccttggctcctgctactgatctgtgctctaccgtgtgaacc120 tgctggaatctctgatgtgagcttgaagacacggcccccaggaggatgggtgatggaggg180 agacaagctggtcctcatctgctcggttgatagagtcactgggaatataacttacttctg240 gtacagaggggccctgggtttccaactggaaacaaagacacaaccttcactaacagcaga300 gtttgagatcagtgacatgaagcagagcgatgctgatcaatattactgtgcggctaacga360 tggccacgaccctatcgccagtgagctggtgagcatccacgtcagagttccagtgtctcg420 ccctgtccttacgtttggggactctggaacccaggctgtgctaggggacctggtggagct480 tcactgtaaggccctgagaggctcacccccaatcttctaccagttttatcatgagagcat540 catcctggggaacagttcagcaccctctggaggaggagcatccttcaacttctccctgac600 tgcagaacattctggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaag660 tgaggtggtggctctcaacttaacaggtctctccttagtgcctactgagaatggaatcag720 ccatctctccttaggactcactgggtggctgcttggctgtcttagccccatcaccatggc780 cttaatattttgctactggctcaagagaaaaataggaagacagtcagaggatccagtcag840 gagccctcctcagactgtgctccaaggatccacgtaccccaaatcccccgactcaaggca900 gccagagcccctgtatgagaacgtgaacgttgtaagtggcaatgaagtgtactctctggt960 gtaccacaccccgcaggtgctggaaccagcagcagctcagcatgtgaggacacacggagt1020 aagtgagtcctttcaggtctcctctggactctattctaagccaaggataaacattgcaca1080 tatggactatgaagacgccatgtagaattatgtaaacagcaactatggagtgctacatac1140 aagcccaaggcctgatgtggcctccaaggatactggggacagggatagcttgccagccca1200 atttccccacacactgcggttcattagatgagtccttcacctaccctgtgtgaagctgga1260 gcaagtcctgcagaaaccacccaggaaaaccaacttagacggagaagccagaagcatttg1320 catctggttgttgcccattcatgttggcacacgaacttttatttacaggaggaaaatggt1380 gtgatgaaagcaactaaggtcttacagcagagggacaatgcgactcagagagcacaaagc1440 cgagatcaatggctttgcaggtctgctgtggagacagagccatgcttcctctgtgcacat1500 accctagagtacttctgagtcactgccatcaacttagaattaaacacagttgcataaaat1560 gtactgt <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>
<223>
Description of Artificial Sequence:/note =

synth etic construct <400>
89 ggctcctgctactgatctgtgctctaccgtgtgaacctgctggaatctct 60 atgctacctt gatgtgagcttgaagacacggcccccaggaggatgggtgatggagggagacaagctggtc 120 ctcatctgctcggttgatagagtcactgggaatataacttacttctggtacagaggggcc 180 ctgggtttccaactggaaacaaagacacaaccttcactaacagcagagtttgagatcagt 240 gacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggccacgaccct 300 atcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccctgtccttacg 360 tttggggactctggaacccaggctgtgctaggggacctggtggagcttcactgtaaggcc 420 ctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatcctggggaac 480 agttcagcaccctctggaggaggagcatccttcaacttctccctgactgcagaacattct 540 ggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgaggtggtggct 600 ctcaacttaacaggtctctccttagtgcctactgagaatggaatcagccatctctcctta 660 ggactcactgggtggctgcttggctgtcttagccccatcaccatggccttaatattttgc 720 tactggctcaagagaaaaataggaagacagtcagaggatccagtcaggagccctcctcag 780 actgtgctccaaggatccacgtaccccaaatcccccgactcaaggcagccagagcccctg 840 tatgagaacgtgaacgttgtaagtggcaatgaagtgtactctctggtgtaccacaccccg 900 caggtgctggaaccagcagcagctcagcatgtgaggacacacggagtaagtgagtccttt 960 caggtctcctctggactctattctaagccaaggataaacattgcacatatggactatgaa 1020 gacgccatgtag <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 90 ggaatctctgatgtgagcttgaagacacggcccccaggaggatgggtgatggagggagac60 aagctggtcctcatctgctcggttgatagagtcactgggaatataacttacttctggtac120 agaggggccctgggtttccaactggaaacaaagacacaaccttcactaacagcagagttt180 gagatcagtgacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggc240 cacgaccctatcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccct300 gtccttacgtttggggactctggaacccaggctgtgctaggggacctggtggagcttcac360 tgtaaggccctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatc420 ctggggaacagttcagcaccctctggaggaggagcatccttcaacttctccctgactgca 480 gaacattctggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgag 540 gtggtggctctcaacttaacaggtctctccttagtgcctactgagaatggaatcagccat 600 ctctccttaggactcactgggtggctgcttggctgtcttagccccatcaccatggcctta 660 atattttgctactggctcaagagaaaaataggaagacagtcagaggatccagtcaggagc 720 cctcctcagactgtgctccaaggatccacgtaccccaaatcccccgactcaaggcagcca 780 gagcccctgtatgagaacgtgaacgttgtaagtggcaatgaagtgtactctctggtgtac 840 cacaccccgcaggtgctggaaccagcagcagctcagcatgtgaggacacacggagtaagt 900 gagtcctttcaggtctcctctggactctattctaagccaaggataaacattgcacatatg 960 gactatgaagacgccatgtag 981 <210>
9l <211>

<212>
DNA

<213> ficial ence Arti Sequ <220>
<223>
Description of Artificial Sequence:/note =

synth etic construct <400>
91 ggctcctgctactgatctgtgctctaccgtgtgaacctgctggaatctct60 atgctacctt gatgtgagcttgaagacacggcccccaggaggatgggtgatggagggagacaagctggtc120 ctcatctgctcggttgatagagtcactgggaatataacttacttctggtacagaggggcc180 ctgggtttccaactggaaacaaagacacaaccttcactaacagcagagtttgagatcagt240 gacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggccacgaccct300 atcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccctgtccttacg360 tttggggactctggaacccaggctgtgctaggggacctggtggagcttcactgtaaggcc420 ctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatcctggggaac480 agttcagcaccctctggaggaggagcatccttcaacttctccctgactgcagaacattct540 ggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgaggtggtggct600 ctcaacttaacaggtctctccttagtgcctactgagaatggaatcagccatctctcctta660 <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>
<223>
Description of Artificial Sequence:/note =

synth etic construct <400>
92 atgtgagcttgaagacacggcccccaggaggatgggtgatggagggagac60 ggaatctctg aagctggtcctcatctgctcggttgatagagtcactgggaatataacttacttctggtac120 agaggggccctgggtttccaactggaaacaaagacacaaccttcactaacagcagagttt180 gagatcagtgacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggc240 cacgaccctatcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccct300 gtccttacgtttggggactctggaacccaggctgtgctaggggacctggtggagcttcac360 tgtaaggccctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatc420 ctggggaacagttcagcaccctctggaggaggagcatccttcaacttctccctgactgca480 gaacattctggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgag540 gtggtggctctcaacttaacaggtctctccttagtgcctactgagaatggaatcagccat600 ctctcctta <210> 93 <211> 303 <212> DNA
<213> Artificial Sequence <220>

<223> Description of Artificial Sequence:/note =

synthetic construct <400> 93 aaaaa taggaagaca gtcagaggatccagtcaggagccctcctcagactgtgctc60 a a g tcaaggcagccagagcccctgtatgagaac120 g a caaggatcca cgtaccccaa atcccccgac gtgaacgttg taagtggcaa tctctggtgtaccacaccccgcaggtgctg180 tgaagtgtac gaaccagcag cagctcagca cacggagtaagtgagtcctttcaggtctcc240 tgtgaggaca tctggactct attctaagcc attgcacatatggactatgaagacgccatg300 aaggataaac tag <210> 94 <211> 1567 <212> DNA

<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
94 ggttttgcttttttttcttttggtgagaggtaccttcaagttaccatccc60 tgagtagtct cagcctggtcctcatgctaccttggctcctgctactgatctgtgctctaccgtgtgaacc120 tgctggaatctctgatgtgagcttgaagacacggcccccaggaggatgggtgatggaggg180 agacaagctggtcctcatctgctcggttgatagagtcactgggaatataacttacttctg240 gtacagaggggccctgggtttccaactggaaacaaagacacaaccttcactaacagcaga300 gtttgagatcagtgacatgaagcagagcgatgctgatcaatattactgtgcggctaacga360 tggccacgaccctatcgccagtgagctggtgagcatccacgtcagagttccagtgtctcg420 ccctgtccttacgtttggggactctggaacccaggctgtgctaggggacctggtggagct480 tcactgtaaggccctgagaggctcacccccaatcttctaccagttttatcatgagagcat540 catcctggggaacagttcagcaccctctggaggaggagcatccttcaacttctccctgac600 tgcagaacattctggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaag660 tgaggtggtggctctcaacttaacaggtctctccttagtgcctactgagaatggaatcag720 ccatctctccttaggactcactgggtggctgcttggctgtcttagccccatcaccatggc780 cttaatattttgctactggctcaagagaaaaataggaagacagtcagaggatccagtcag840 gagccctcctcagactgtgctccaaggatccacgtaccccaaatcccccgactcaaggca900 gccagagcccctgtatgagaacgtgaacgttgtaagtggcaatgaagtgtactctctggt960 gtaccacaccccgcaggtgctggaaccagcagcagctcagcatgtgaggacacacggagt1020 aagtgagtcctttcaggtctcctctggactctattctaagccaaggataaacattgcaca1080 tatggactatgaagacgccatgtagaattatgtaaacagcaactatggagtgctacatac1140 aagcccaaggcctgatgtggcctccaaggatactggggacagggatagcttgccagccca1200 atttccccacacactgcggttcattagatgagtccttcacctaccctgtgtgaagctgga1260 gcaagtcctgcagaaaccacccaggaaaaccaacttagacggagaagccagaagcatttg1320 catctggttgttgcccattcatgttggcacacgaacttttatttacaggaggaaaatggt1380 gtgatgaaagcaactaaggtcttacagcagagggacaatgcgactcagagagcacaaagc1440 cgagatcaatggctttgcaggtctgctgtggagacagagccatgcttcctctgtgcacat1500 accctagagtacttctgagtcactgccatcaacttagaattaaacacagttgcataaaat1560 gtactgt <210> 95 <211> 903 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 95 atgctacctt ggctcctgct actgatctgt gctctaccgt gtgaacctgc tggaatctct 60 gatgtgagct tgaagacacg gcccccagga ggatgggtga tggagggaga caagctggtc 120 ctcatctgctcggttgatagagtcactgggaatataacttacttctggtacagaggggcc180 ctgggtttccaactggaaacaaagacacaaccttcactaacagcagagtttgagatcagt240 gacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggccacgaccct300 atcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccctgtccttacg360 tttggggactctggaacccaggctgtgctaggggacctggtggagcttcactgtaaggcc920 ctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatcctggggaac480 agttcagcaccctctggaggaggagcatccttcaacttctccctgactgcagaacattct540 ggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgaggtggtggct600 ctcaacttaacaggaagacagtcagaggatccagtcaggagccctcctcagactgtgctc660 caaggatccacgtaccccaaatcccccgactcaaggcagccagagcccctgtatgagaac720 gtgaacgttgtaagtggcaatgaagtgtactctctggtgtaccacaccccgcaggtgctg780 gaaccagcagcagctcagcatgtgaggacacacggagtaagtgagtcctttcaggtctcc840 tctggactctattctaagccaaggataaacattgcacatatggactatgaagacgccatg900 tag <2l0> 96 <211> 852 <212> DNA
<213> Artificial Sequence <220>

<223> equence:/note =
Description of Artificial S

synthetic construct <400>
96 atgtgagcttgaagacacggcccccaggaggatgggtgatggagggagac60 ggaatctctg aagctggtcctcatctgctcggttgatagagtcactgggaatataacttacttctggtac120 agaggggccctgggtttccaactggaaacaaagacacaaccttcactaacagcagagttt180 gagatcagtgacatgaagcagagcgatgctgatcaatattactgtgcggctaacgatggc240 cacgaccctatcgccagtgagctggtgagcatccacgtcagagttccagtgtctcgccct300 gtccttacgtttggggactctggaacccaggctgtgctaggggacctggtggagcttcac360 tgtaaggccctgagaggctcacccccaatcttctaccagttttatcatgagagcatcatc420 ctggggaacagttcagcaccctctggaggaggagcatccttcaacttctccctgactgca480 gaacattctggaaacttctcctgtgaggccagcaatggacagggtgcccaacgaagtgag540 gtggtggctctcaacttaacaggaagacagtcagaggatccagtcaggagccctcctcag600 actgtgctccaaggatccacgtaccccaaatcccccgactcaaggcagccagagcccctg660 tatgagaacgtgaacgttgtaagtggcaatgaagtgtactctctggtgtaccacaccccg720 caggtgctggaaccagcagcagctcagcatgtgaggacacacggagtaagtgagtccttt780 caggtctcctctggactctattctaagccaaggataaacattgcacatatggactatgaa840 gacgccatgtag <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>

<223>
Description of Artificial Sequence:/note =

synthetic construct <400>
97 cactcaactgttttagaagagcagttccccagatttctccttggagctgt60 attcaagtta gagtgactaccattgcgagcaagagcaagaggaaagcactacctgtgagcagatgtctgg120 ttcattctcaccctgtgtggtgttcacacagatgtggctgactctactggttgtgactcc180 tgtcaatggacagcatgaagctgcacagcagtctgtggtttcccttcagcctccatggac240 cactttctttcgaggagaggtcgtcacactgacttgttatagattcggcttctccgtacc300 ccagaaaacaaaatggtaccagaaaagaaaaacagtgaagcaaaccccaggtgctttggt360 aattaaagcacataccttaaaggtccatgagtccggagagtattggtgccaagccgacag420 cttacttccgagcatgcacgtgaacgtagagttttctgaagattttctggtgctgcaagc480 tccacctgctgtgtttgaaggagactctgtggttctgaggtgctacgcaaagaaaggcat540 agaagcagagaccctgacattttacaaggatggtaaagctctgacattacatcatcaaag600 tgagctctctattcatcatgcaaatctgaaggacaacggtcaatacaaatgcacttcgaa660 gaagaagtggtcttttgggtccctctatacttccaatacggtcggagttcaagtccaaga720 gttgttcccacggcctgtgctgagagccagaccctcccatcccatagatggaagtccagt780 gaccctgacgtgtcagacccagctctctgcacagaagtcagatgcccggctccagttctg840 tttcttcagaaacctccagcttctggggtcaggctgcagccgctcctcagagtttcacat900 tcctgccatatggactgaagagtcaaggagataccagtgcaaggcagaaacagtgaattc960 ccaagttagaaaacaaagtacagcgttcataatcccagtgcagagagcttctgcgagatt1020 ccaaacacacatcatcccagcctcaaagttggtgtttgaagggcagttgctgttactcaa1080 ctgctcagtaaaaggagtyccaggrcccctcaaattctcctggtataaaaaggacatgct1140 gaatgaagaaacaaagattcttaagtcctccaacgcagaattcaagatctcccaggtgaa1200 catcagtgacgcaggggagtatcactgtgaagctaccaacagccgccgaagctttgtcag1260 cagggcatttcccatcaccataaaagtcccagtatctcaaccagttctcaccctaagcac1320 aggcaagacccaggcccttgagggagacttgatgacacttcattgtcaatcccagagggg1380 ctctccatgtatcctgtatgaattcttctatgagaatgtctccctggggaatagctctat1440 actctctggaggaggagcatacttcaatttctctatgagcacagagcgatctggaaacta1500 ctactgcacagcagacaatggcctgggagcccagtgcagtgaagctataaggatctctat1560 ctttgacatgacaaagaacagaagtgttcctatggctgccggaatcactgtgggactgct1620 catcatggctgttggagtgtttctgttttattgctggttctctagaaaagcaggaggaaa1680 gcctacctctgatgactccagaaacccttcagattcagaaccccaggagcccacctatta1740 caacgtaccagcctgtatagaactgcagccagtgtacagcaatgagcctgaggaaaacgt1800 gatttacacagaagtacggagaactcaaccaagacagaaacatgcagatcaggagtctga1860 aagcccaagatcaaggtgccagatggctgagaaaaagtaggatatgtctcctccaagaac1920 agctccagaaaagaaacccgaagcttcgtcagtctaatctcaccgatgcttctactgggc1980 ctgcactttcctacccacggatggctccacagatcatggacagcaaggaaatggccaact2040 ctcctaagactgggccaacatccccatcttctctttggtttcccagagccacgccacccc2100 aaagtcagcaggaagttgcaaaagatcacaacgaccctattcctgttttgtaaccacccc2160 cagcctgaagcaggctgagccagaccttgaccttgctgccactaaggagattacctaggg2220 tggagcctgcctctctagatcactctattgttcagccactgccactgttctccttcaaga2280 cactgctacctgctgggaggccactgagctattccagagactacaccctatcctgcacat2340 catcacctgtagcctgttccaggctccaagaatgaattggcggcaatgggcctcccccct2400 accccctttataagtgcatttgccattaaacatttgggctttgatct 2447 <210> 98 <211> 1788 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
98 cattctcaccctgtgtggtgttcacacagatgtggctgactctactggtt60 atgtctggtt gtgactcctgtcaatggacagcatgaagctgcacagcagtctgtggtttcccttcagcct120 ccatggaccactttctttcgaggagaggtcgtcacactgacttgttatagattcggcttc180 tccgtaccccagaaaacaaaatggtaccagaaaagaaaaacagtgaagcaaaccccaggt240 gctttggtaattaaagcacataccttaaaggtccatgagtccggagagtattggtgccaa300 gccgacagcttacttccgagcatgcacgtgaacgtagagttttctgaagattttctggtg360 ctgcaagctccacctgctgtgtttgaaggagactctgtggttctgaggtgctacgcaaag420 aaaggcatagaagcagagaccctgacattttacaaggatggtaaagctctgacattacat480 catcaaagtgagctctctattcatcatgcaaatctgaaggacaacggtcaatacaaatgc540 acttcgaagaagaagtggtcttttgggtccctctatacttccaatacggtcggagttcaa600 gtccaagagttgttcccacggcctgtgctgagagccagaccctcccatcccatagatgga660 agtccagtgaccctgacgtgtcagacccagctctctgcacagaagtcagatgcccggctc720 cagttctgtttcttcagaaacctccagcttctggggtcaggctgcagccgctcctcagag780 tttcacattcctgccatatggactgaagagtcaaggagataccagtgcaaggcagaaaca840 gtgaattcccaagttagaaaacaaagtacagcgttcataatcccagtgcagagagcttct900 gcgagattccaaacacacatcatcccagcctcaaagttggtgtttgaagggcagttgctg960 ttactcaactgctcagtaaaaggagtyccaggrcccctcaaattctcctggtataaaaag1020 gacatgctgaatgaagaaacaaagattcttaagtcctccaacgcagaattcaagatctcc1080 caggtgaacatcagtgacgcaggggagtatcactgtgaagctaccaacagccgccgaagc1140 tttgtcagcagggcatttcccatcaccataaaagtcccagtatctcaaccagttctcacc1200 ctaagcacaggcaagacccaggcccttgagggagacttgatgacacttcattgtcaatcc1260 cagaggggctctccatgtatcctgtatgaattcttctatgagaatgtctccctggggaat1320 agctctatactctctggaggaggagcatacttcaatttctctatgagcacagagcgatct1380 ggaaactactactgcacagcagacaatggcctgggagcccagtgcagtgaagctataagg1440 atctctatctttgacatgacaaagaacagaagtgttcctatggctgccggaatcactgtg1500 ggactgctcatcatggctgttggagtgtttctgttttattgctggttctctagaaaagca1560 ggaggaaagcctacctctgatgactccagaaacccttcagattcagaaccccaggagccc1620 acctattacaacgtaccagcctgtatagaactgcagccagtgtacagcaatgagcctgag1680 gaaaacgtgatttacacagaagtacggagaactcaaccaagacagaaacatgcagatcag1740 gagtctgaaagcccaagatcaaggtgccagatggctgagaaaaagtag 1788 <210> 99 <211> 1710 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
99 ctgcacagcagtctgtggtttcccttcagcctccatggaccactttcttt60 cagcatgaag cgaggagaggtcgtcacactgacttgttatagattcggcttctccgtaccccagaaaaca120 aaatggtaccagaaaagaaaaacagtgaagcaaaccccaggtgctttggtaattaaagca180 cataccttaaaggtccatgagtccggagagtattggtgccaagccgacagcttacttccg240 agcatgcacgtgaacgtagagttttctgaagattttctggtgctgcaagctccacctgct300 gtgtttgaaggagactctgtggttctgaggtgctacgcaaagaaaggcatagaagcagag360 accctgacattttacaaggatggtaaagctctgacattacatcatcaaagtgagctctct420 attcatcatgcaaatctgaaggacaacggtcaatacaaatgcacttcgaagaagaagtgg480 tcttttgggtccctctatacttccaatacggtcggagttcaagtccaagagttgttccca540 cggcctgtgctgagagccagaccctcccatcccatagatggaagtccagtgaccctgacg600 tgtcagacccagctctctgcacagaagtcagatgcccggctccagttctgtttcttcaga660 aacctccagcttctggggtcaggctgcagccgctcctcagagtttcacattcctgccata720 tggactgaagagtcaaggagataccagtgcaaggcagaaacagtgaattcccaagttaga780 aaacaaagtaoagcgttcataatcccagtgcagagagcttctgcgagattccaaacacac840 atcatcccagcctcaaagttggtgtttgaagggcagttgctgttactcaactgctcagta900 aaaggagtyccaggrcccctcaaattctcctggtataaaaaggacatgctgaatgaagaa960 acaaagattcttaagtcctccaacgcagaattcaagatctcccaggtgaacatcagtgac1020 gcaggggagtatcactgtgaagctaccaacagccgccgaagctttgtcagcagggcattt1080 cccatcaccataaaagtcccagtatctcaaccagttctcaccctaagcacaggcaagacc1140 caggcccttgagggagacttgatgacacttcattgtcaatcccagaggggctctccatgt1200 atcctgtatgaattcttctatgagaatgtctccctggggaatagctctatactctctgga1260 ggaggagcatacttcaatttctctatgagcacagagcgatctggaaactactactgcaca1320 gcagacaatggcctgggagcccagtgcagtgaagctataaggatctctatctttgacatg1380 acaaagaacagaagtgttcctatggctgccggaatcactgtgggactgctcatcatggct1440 gttggagtgtttctgttttattgctggttctctagaaaagcaggaggaaagcctacctct1500 gatgactccagaaacccttcagattcagaaccccaggagcccacctattacaacgtacca1560 gcctgtatagaactgcagccagtgtacagcaatgagcctgaggaaaacgtgatttacaca1620 gaagtacggagaactcaaccaagacagaaacatgcagatcaggagtctgaaagcccaaga1680 tcaaggtgccagatggctgagaaaaagtag 1710 <210> 100 <211> 1401 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400>
100 ctgcacagcagtctgtggtttcccttcagcctccatggaccactttcttt60 cagcatgaag cgaggagaggtcgtcacactgacttgttatagattcggcttctccgtaccccagaaaaca120 aaatggtaccagaaaagaaaaacagtgaagcaaaccccaggtgctttggtaattaaagca180 cataccttaaaggtccatgagtccggagagtattggtgccaagccgacagcttacttccg240 agcatgcacgtgaacgtagagttttctgaagattttctggtgctgcaagctccacctgct300 gtgtttgaaggagactctgtggttctgaggtgctacgcaaagaaaggcatagaagcagag360 accctgacattttacaaggatggtaaagctctgacattacatcatcaaagtgagctctct420 attcatcatgcaaatctgaaggacaacggtcaatacaaatgcacttcgaagaagaagtgg480 tcttttgggtccctctatacttccaatacggtcggagttcaagtccaagagttgttccca540 cggcctgtgctgagagccagaccctcccatcccatagatggaagtccagtgaccctgacg600 tgtcagacccagctctctgcacagaagtcagatgcccggctccagttctgtttcttcaga660 aacctccagcttctggggtcaggctgcagccgctcctcagagtttcacattcctgccata720 tggactgaagagtcaaggagataccagtgcaaggcagaaacagtgaattcccaagttaga780 aaacaaagtacagcgttcataatcccagtgcagagagcttctgcgagattccaaacacac840 atcatcccagcctcaaagttggtgtttgaagggcagttgctgttactcaactgctcagta900 aaaggagtyccaggrcccctcaaattctcctggtataaaaaggacatgctgaatgaagaa960 acaaagattcttaagtcctccaacgcagaattcaagatctcccaggtgaacatcagtgac1020 gcaggggagtatcactgtgaagctaccaacagccgccgaagctttgtcagcagggcattt1080 cccatcaccataaaagtcccagtatctcaaccagttctcaccctaagcacaggcaagacc1140 caggcccttgagggagacttgatgacacttcattgtcaatcccagaggggctctccatgt1200 atcctgtatgaattcttctatgagaatgtctccctggggaatagctctatactctctgga1260 ggaggagcatacttcaatttctctatgagcacagagcgatctggaaactactactgcaca1320 gcagacaatggcctgggagcccagtgcagtgaagctataaggatctctatctttgacatg1380 acaaagaacagaagtgttcct 1401 <210>

<211>

<212>
DNA

<213> ficial ence Arti Sequ <220>
<223> Description of Artificial Sequence:/note =

synth etic construct <400> 101 atgtctggttcattctcaccctgtgtggtgttcacacagatgtggctgactctactggtt 60 gtgactcctgtcaatggacagcatgaagctgcacagcagtctgtggtttcccttcagcct 120 ccatggaccactttctttcgaggagaggtcgtcacactgacttgttatagattcggcttc 180 tccgtaccccagaaaacaaaatggtaccagaaaagaaaaacagtgaagcaaaccccaggt 240 gctttggtaattaaagcacataccttaaaggtccatgagtccggagagtattggtgccaa 300 gccgacagcttacttccgagcatgcacgtgaacgtagagttttctgaagattttctggtg 360 ctgcaagctccacctgctgtgtttgaaggagactctgtggttctgaggtgctacgcaaag 420 aaaggcatagaagcagagaccctgacattttacaaggatggtaaagctctgacattacat 980 catcaaagtgagctctctattcatcatgcaaatctgaaggacaacggtcaatacaaatgc 540 acttcgaagaagaagtggtcttttgggtccctctatacttccaatacggtcggagttcaa 600 gtccaagagttgttcccacggcctgtgctgagagccagaccctcccatcccatagatgga 660 agtccagtgaccctgacgtgtcagacccagctctctgcacagaagtcagatgcccggctc 720 cagttctgtttcttcagaaacctccagcttctggggtcaggctgcagccgctcctcagag 780 tttcacattcctgccatatggactgaagagtcaaggagataccagtgcaaggcagaaaca 840 gtgaattcccaagttagaaaacaaagtacagcgttcataatcccagtgcagagagcttct 900 gcgagattccaaacacacatcatcccagcctcaaagttggtgtttgaagggcagttgctg 960 ttactcaactgctcagtaaaaggagtyccaggrcccctcaaattctcctggtataaaaag 1020 gacatgctgaatgaagaaacaaagattcttaagtcctccaacgcagaattcaagatctcc 1080 caggtgaacatcagtgacgcaggggagtatcactgtgaagctaccaacagccgccgaagc 1140 tttgtcagcagggcatttcccatcaccataaaagtcccagtatctcaaccagttctcacc 1200 ctaagcacaggcaagacccaggcccttgagggagacttgatgacacttcattgtcaatcc 1260 cagaggggctctccatgtatcctgtatgaattcttctatgagaatgtctccctggggaat 1320 agctctatactctctggaggaggagcatacttcaatttctctatgagcacagagcgatct 1380 ggaaactactactgcacagcagacaatggcctgggagcccagtgcagtgaagctataagg 1440 atctctatctttgacatgacaaagaacagaagtgttcct 1479 <210> 102 <211> 240 <212> DNA
<213> Artificial Sequence <220>
<223> Description of Artificial Sequence:/note =
synthetic construct <400> 102 tctagaaaag caggaggaaa gcctacctct gatgactcca gaaacccttc agattcagaa 60 ccccaggagc ccacctatta caacgtacca gcctgtatag aactgcagcc agtgtacagc 120 aatgagcctg aggaaaacgt gatttacaca gaagtacgga gaactcaacc aagacagaaa 180 catgcagatc aggagtctga aagcccaaga tcaaggtgcc agatggctga gaaaaagtag 240

Claims (219)

What is claimed is:

1. An isolated FcRH, comprising a cytoplasmic region with more than 107 or less than 104 amino acids, a transmembrance region, and an extracellular region.

2. The isolated FcRH of claim 1, wherein the extracellular region comprises less than four Ig domains.

3. The isolated FcRH of claim 2, wherein the cytoplamic region comprises less than 104 amino acids.

4. The isolated FcRH of claim 3, wherein the transmembrane region comprises an acidic amino acid.

5. The isolated FcRH of claim 4, wherein the acidic amino acid is glutamate.

6. The isolated FcRH of claim 2, wherein the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:1

7. The isolated FcRH of claim 2, wherein the extracellular region comprises the amino acid sequence of SEQ ID NO:21.

8. The isolated FcRH of claim 1, comprising the amino acid sequence of SEQ ID
NO:2.

9. The isolated FcRH of claim 1, wherein the receptor is expressed by myeloid cells.

10. The isolated FcRH of claim 9, wherein the receptor is expressed by T-cells.

11. A polypeptide comprising the amino acid sequence of SEQ ID NO: 1.

12. A polypeptide comprising the amino acid of SEQ ID NO:1 with conservative amino acid substitutions.

13. A polypeptide comprising the amino acid sequence of SEQ ID NO:21

14. A polypeptide comprising the amino acid of SEQ ID NO:21 with conservative amino acid substitutions.

15. A polypeptide comprising the amino acid of SEQ ID NO:2.

16. A polypeptide comprising the amino acid of SEQ ID NO:2 with conservative amino acid substitutions.

17. The isolated FcRH of claim 1, wherein the cytoplasmic region comprises less than 99 amino acids and wherein the receptor further comprises an extracellular region with up to four Ig domain and up to five N-linked glycosylation sites.

18. The isolated FcRH of claim 17, wherein the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:3.

19. The isolated FcRH of claim 17, wherein the extracellular region comprises the amino acid sequence of SEQ ID NO:22.

20. The isolated FcRH of claim 1, comprising the amino acid sequence of SEQ ID
NO:4.

21. A polypeptide comprising the amino acid sequence of SEQ ID NO:3.

22. A polypeptide comprising the amino acid of SEQ ID NO:3 with conservative amino acid substitutions.

23. A polypeptide comprising the amino acid sequence of SEQ ID NO:22

24. A polypeptide comprising the amino acid of SEQ ID NO:22 with conservative amino acid substitutions.

25. A polypeptide comprising the amino acid of SEQ ID NO:4.

26. A polypeptide comprising the amino acid of SEQ ID NO:4 with conservative amino acid substitutions.

27. The isolated FcRH of claim 1, wherein the cytoplasmic region comprises more than 107 amino acids.

28. The isolated FcRH of claim 27, wherein the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:5.

29. The isolated FcRH of claim 27, wherein the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:23.

30. The isolated FcRH of claim 27, wherein the extracellular region comprises the amino acid sequence of SEQ ID NO:24.

31. The isolated FcRH of claim 1, comprising the amino acid sequence of SEQ ID
NO:6.

32. The isolated FcRH of claim 1, comprising the amino acid sequence of SEQ ID
NO:25.

33. A polypeptide comprising the amino acid sequence of SEQ ID NO:5

34. A polypeptide comprising the amino acid of SEQ ID NO:5 with conservative amino acid substitutions.

35. A polypeptide comprising the amino acid sequence of SEQ ID NO:24

36. A polypeptide comprising the amino acid of SEQ ID NO:24 with conservative amino acid substitutions.

37. A polypeptide comprising the amino acid sequence of SEQ ID NO:23

38. A polypeptide comprising the amino acid of SEQ ID NO:23 with conservative amino acid substitutions.

39. A polypeptide comprising the amino acid sequence of SEQ ID NO:6.

40. A polypeptide comprising the amino acid of SEQ ID NO:6 with conservative amino acid substitutions.

41. A polypeptide comprising the amino acid sequence of SEQ ID NO:25.

42. A polypeptide comprising the amino acid of SEQ ID NO:25 with conservative amino acid substitutions.

43. The isolated FcRH of claim 1, wherein the cytoplasmic region comprises the amino acid sequence of SEQ ID NO:26.

44. The isolated FcRH of claim 1, wherein the extracellular region comprises the amino acid sequence of SEQ ID NO:27.

45. The isolated FcRH of claim 1, comprising the amino acid sequence of SEQ ID
NO:28.

46. An isolated nucleic acid, comprising a nucleotide sequence that encodes the FcRH
of claim 2.

47. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:1.

48. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:21.

49. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:2.

50. The nucleic acid of claim 46, comprising the nucleotide sequence of SEQ ID
NO:7.

51. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:7, or the complement of SEQ ID
NO:7.

52. The nucleic acid of claim 46, comprising the nucleotide sequence of SEQ ID
NO:13.

53. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:13 or the complement of SEQ
ID NO:13.

54. The nucleic acid of claim 46, comprising the nucleotide sequence of SEQ ID
NO:8.

55. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:8, or the complement of SEQ ID
NO:8.

56. A single stranded nucleic acid that hybridizes under stringent conditions to a nucleic acid having the sequence of SEQ ID NO:7, SEQ ID NO:13 or SEQ ID
NO:8.

57. An isolated nucleic acid, comprising a nucleotide sequence that encodes the FcRH
of claim 17.

58. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:3.

59. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:22.

60. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:4.

61. The nucleic acid of claim 57, comprising the nucleotide sequence of SEQ ID
NO:9.

62. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:9, or the complement of SEQ ID
NO:9.

63. The nucleic acid of claim 57, comprising the nucleotide sequence of SEQ ID
NO:14.

64. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:14, or the complement of SEQ
ID NO:14.

65. The nucleic acid of claim 57, comprising the nucleotide sequence of SEQ ID
NO:10.

66. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:10, or the complement of SEQ
ID NO:10.

67. A single stranded nucleic acid that hybridizes under stringent conditions to a nucleic acid having the sequence of SEQ ID NO:9, SEQ ID NO:14, or SEQ ID
NO:10.

68. An isolated nucleic acid, comprising a nucleotide sequence that encodes the FcRH
of claim 27.

69. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:5.

70. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:23.

71. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:24.

72. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:6.

73. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:25.

74. The nucleic acid of claim 68, comprising the nucleotide sequence of SEQ ID
NO:11.

75. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:11, or the complement of SEQ
ID NO:11.

76. The nucleic acid of claim 68, comprising the nucleotide sequence of SEQ ID
NO:16.

77. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:16, or the complement of SEQ
ID NO:16.

78. The nucleic acid of claim 68, comprising the nucleotide sequence of SEQ ID
NO:15.

79. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:15, or the complement of SEQ
ID NO:15.

80. The nucleic acid of claim 68, comprising the nucleotide sequence of SEQ ID
NO:12.

81. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:12, or the complement of SEQ
ID NO:12.

82. The nucleic acid of claim 68, comprising the nucleotide sequence of SEQ ID
NO:17.

83. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:11, or the complement of SEQ
ID NO:17.

84. A single stranded nucleic acid that hybridizes under stringent conditions to a nucleic acid having the sequence of SEQ ID NO:11, SEQ ID NO:15, SEQ ID
NO:16, or SEQ ID NO:12.

85. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:26.

86. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:27.

87. An isolated nucleic acid, comprising a nucleotide sequence that encodes SEQ ID
NO:28.

88. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:18, or the complement of SEQ
ID NO:18.

89. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:19, or the complement of SEQ
ID NO:19.

90. An isolated nucleic acid comprising a sequence that hybridizes under highly stringent conditions to a hybridization probe, wherein the hybridization probe comprises the nucleotide sequence of SEQ ID NO:20, or the complement of SEQ
ID NO:20.

91. A single stranded nucleic acid that hybridizes under stringent conditions to a nucleic acid having the sequence of SEQ ID NO:18, SEQ ID NO:19, or SEQ ID
NO:20.

92. An expression vector comprising the nucleic acid of claim 46 operably linked to an expression control sequence.

93. An isolated cell comprising the vector of claim 92.

94. A method of making a FcRH, comprising culturing the cell of claim 93 under conditions permitting expression of the FcRH.

95. An expression vector comprising the nucleic acid of claim 57 operably linked to an expression control sequence.

96. An isolated cell comprising the vector of claim 95.

97. A method of making a FcRH, comprising culturing the cell of claim 96 under conditions permitting expression of the FcRH.

98. An expression vector comprising the nucleic acid of claim 56 operably linked to an expression control sequence.

99. An isolated cell comprising the vector of claim 98.

100. A method of making a FcRH, comprising culturing the cell of claim 99 under conditions permitting expression of the FcRH.

101. An expression vector comprising the nucleic acid of claim 57 operably linked to an expression control sequence.

102. An isolated cell comprising the vector of claim 101.

103. A method of making a FcRH, comprising culturing the cell of claim 102 under conditions permitting expression of the FcRH.

104. An expression vector comprising the nucleic acid of claim 67 operably linked to an expression control sequence.

105. An isolated cell comprising the vector of claim 104.

106. A method of making a FcRH, comprising culturing the cell of claim 105 under conditions permitting expression of the FcRH.

107. An expression vector comprising the nucleic acid of claim 68 operably linked to an expression control sequence.

108. An isolated cell comprising the vector of claim 107.

109. A method of making a FcRH, comprising culturing the cell of claim 108 under conditions permitting expression of the FcRH.

110. An expression vector comprising the nucleic acid of claim 91 operably linked to an expression control sequence.

111. An isolated cell comprising the vector of claim 110.

112. A method of making a FcRH, comprising culturing the cell of claim 111 under conditions permitting expression of the FcRH.

113. A purified antibody or immunonologic fragment thereof, wherein the antibody or fragment thereof selectively binds to the FcRH of claim 1.

114. A purified antibody or immunonologic fragment thereof, wherein the antibody or fragment thereof selectively binds to the FcRH of claim 2.

115. A purified antibody or immunonologic fragment thereof, wherein the antibody or fragment thereof selectively binds to the FcRH of claim 17.

116. A purified antibody or immunonologic fragment thereof, wherein the antibody or fragment thereof selectively binds to the FcRH of claim 27.

117. The antibody or fragment of claim 113, wherein the antibody or fragment is a monoclonal antibody or fragment thereof.

118. The antibody or fragment of claim 113, wherein the antibody or fragment thereof is a humanized antibody, a fully human antibody, or a fragment thereof.

119. The antibody or fragment of claim 113, wherein the antibody or fragment thereof is a single chain antibody or fragment thereof.

120. The antibody or fragment of claim 113, wherein the antibody or fragment thereof is labeled.

121. The antibody or fragment of claim 113, wherein the label is a radiolabel.

122. The antibody or fragment of claim 113, wherein the antibody or fragment is conjugated or fused with a toxin.

123. A purified antibody that selectively binds to the FcRH of claim 6, but not to the FcRH of claim 18, 28, or 43.

124. A purified antibody that selectively binds to the FcRH of claim 18, but not to the FcRH of claim 6, 28, or 43.

125. A purified antibody that selectively binds to the FcRH of claim 28, but not to the FcRH of claim 6, 18, or 43.

126. The purified antibody of claim 125, wherein the antibody does not bind to the FcRH of claim 29.

127. A purified antibody that selectively binds to the FcRH of claim 29, but not to the FcRH of claim 6, 18, or 43.

128. The purified antibody of claim 127, wherein the antibody does not bind to the FcRH of claim 28.

129. A purified antibody that selectively binds to the FcRH of claim 43, but not to the FcRH of claim 6, 18, or 28.

130. A purified antibody that selectively binds to the FcRH of claim 7, but not to the FcRH of claim 19, 30, or 44.

131. A purified antibody that selectively binds to the FcRH of claim 19, but not to the FcRH of claim 7, 30, or 44.

132. A purified antibody that selectively binds to the FcRH of claim 30, but not to the FcRH of claim 7, 19, or 44.

133. A purified antibody that selectively binds to the FcRH of claim 44, but not to the FcRH of claim 7, 19, or 30.

134. A method of diagnosing a malignancy of hematopoietic cell lineage in a subject, comprising:
(a) contacting a biological sample of the subject with the antibody of claim 113 under conditions that allow the antibody to bind to an FcRH in the biological sample;
(b) detecting the amount or pattern of binding by the antibody, changes in the amount or pattern of binding as compared to binding in a control sample indicating a malignancy of hematopoietic cell lineage in the subject.

135. The method of claim 134, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

136. The method of claim 134, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

137. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:1.

138. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:21.

139. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:2.

140. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:3.

141. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:22.

142. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:4.

143. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:5.

144. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:23.

145. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:24.

146. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:6.

147. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:25.

148. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:26.

149. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:27.

150. The method of claim 134, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:28.

151. A method of diagnosing a malignancy of hematopoietic cell lineage in a subject, comprising:
(a) contacting the nucleic acid of claim 56 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with an FcRH in the biological sample;

(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating a malignancy of hematopoietic cell lineage in the subject.

152. The method of claim 151, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

153. The method of claim 151, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

154. A method of diagnosing a malignancy of hematopoietic cell lineage in a subject, comprising:
(a) contacting the nucleic acid of claim 67 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating a malignancy of hematopoietic cell lineage.

155. The method of claim 154, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

156. The method of claim 154, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

157. A method of diagnosing a malignancy of hematopoietic cell lineage in a subject, comprising:
(a) contacting the nucleic acid of claim 84 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating a malignancy of hematopoietic cell lineage.

158. The method of claim 157, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

159. The method of claim 157, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

160. A method of diagnosing a malignancy of hematopoietic cell lineage in a subject, comprising:
(a) contacting the nucleic acid of claim 91 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating a malignancy of hematopoietic cell lineage.

161. The method of claim 160, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

162. The method of claim 160, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

163. A method of treating a malignancy of hematopoietic cell lineage in a subject, comprising contacting the subject's malignant cells with a therapeutically effective amount of the antibody of claim 113.

164. The method of claim 163, wherein the malignancy of hematopoietic cell lineage is a malignancy of B cell lineage.

165. The method of claim 163, wherein the malignancy of hematopoietic cell lineage is a malignancy of T cell lineage.

166. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:1.

167. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:21.

168. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:2.

169. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:3.

170. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:22.

171. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:4.

172. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:5.

173. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:23.

174. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:24.

175. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:6.

176. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:25.

177. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:26.

178. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:27.

179. The method of claim 163, wherein the antibody selectively binds an FcRH
having the amino acid sequence of SEQ ID NO:28.

180. A method of treating a malignancy of hematopoietic cell lineage in a subject, comprising contacting the subject's malignant cells with a therapeutically effective amount of the nucleic acid of claim 56.

181. A method of treating a malignancy of hematopoietic cell lineage in a subject, comprising contacting the subject's malignant cells with a therapeutically effective amount of the nucleic acid of claim 67.

182. A method of treating a malignancy of hematopoietic cell lineage in a subject, comprising contacting the subject's malignant cells with a therapeutically effective amount of the nucleic acid of claim 84.

183. A method of treating a malignancy of hematopoietic cell lineage in a subject, comprising contacting the subject's malignant cells with a therapeutically effective amount of the nucleic acid of claim 91.

184. A method of diagnosing an autoimmune disease in a subject, comprising:
(a) contacting a biological sample of the subject with the antibody of claim 113 under conditions that allow the antibody to bind to FcRH in the biological sample;
(b) detecting the amount or pattern of binding by the antibody, changes in the amount or pattern of binding as compared to binding in a control sample indicating an autoimmune disease in the subject.

185. A method of diagnosing an autoimmune disease in a subject, comprising:
(a) contacting the nucleic acid of claim 56 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating an autoimmune disease.

186. A method of diagnosing an autoimmune disease in a subject, comprising:
(a) contacting the nucleic acid of claim 67 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating an autoimmune disease.

187. A method of diagnosing an autoimmune disease in a subject, comprising:
(a) contacting the nucleic acid of claim 84 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating an autoimmune disease.

188. A method of diagnosing an autoimmune disease in a subject, comprising:
(a) contacting the nucleic acid of claim 91 with a biological sample of the subject under conditions that allow the nucleic acid to hybridize with the biological sample;
(b) detecting the amount or pattern of binding by the nucleic acid, changes in the amount or pattern of binding as compared to binding in a control sample indicating an autoimmune disease.

189. A method of treating an autoimmune disease in a subject, comprising contacting, with a therapeutically effective amount of the antibody of claim 113, one or more FcRH expressing cells of the subject.

190. A method of treating an autoimmune disease in a subject, comprising contacting, with a therapeutically effective amount of the nucleic acid of claim 56, FcRH expressing cells of the subject.

191. A method of treating an autoimmune disease in a subject, comprising contacting, with a therapeutically effective amount of the nucleic acid of claim 67, FcRH expressing cells of the subject.

192. A method of treating an autoimmune disease in a subject, comprising contacting, with a therapeutically effective amount of the nucleic acid of claim 84, FcRH expressing cells of the subject.

193. A method of treating an autoimmune disease in a subject, comprising contacting, with a therapeutically effective amount of the nucleic acid of claim 91, FcRH expressing cells of the subject.

194. A method of modulating a humoral immune response in a subject, comprising administering to the subject the isolated FcRH of claim 1.

195. A method of modulating a humoral immune response in a subject, comprising administering to the subject the antibody of claim 113.

196. A method of modulating a humoral immune response in a subject, comprising administering to the subject the nucleic acid of claim 56.

197. A method of modulating a humoral immune response in a subject, comprising administering to the subject the nucleic acid of claim 67.

198. A method of modulating a humoral immune response in a subject, comprising administering to the subject the nucleic acid of claim 84.

199. A method of modulating a humoral immune response in a subject, comprising administering to the subject the nucleic acid of claim 91.

200. An isolated mouse FcRH isoform of FcRH1, wherein the isoform lacks a cytoplasmic region.

201. A polypeptide comprising the amino acid sequence of SEQ ID NO:70.

202. A polypeptide comprising the amino acid of SEQ ID NO:70 with conservative amino acid substitutions.

203. An isolated mouse FcRH isoform of FcRH2, wherein the FcRH lacks a transmembrane region.

204. A polypeptide comprising the amino acid sequence of SEQ ID NO:73.

205. A polypeptide comprising the amino acid of SEQ ID NO:73 with conservative amino acid substitutions.

206. A polypeptide comprising the amino acid of SEQ ID NO:.77.

207. A polypeptide comprising the amino acid of SEQ ID NO:77 with conservative amino acid substitutions.

208. A polypeptide comprising the amino acid sequence of SEQ ID NO:78.

209. A polypeptide comprising the amino acid sequence of SEQ ID NO:78 with conservative amino acid substitutions.

210. A nucleic acid encoding the isolated mouse FcRH isoform of claim 200.

211. A nucleic acid encoding the isolated mouse FcRH isoform of claim 203.

212. A nucleic acid encoding the polypeptide of claim 201.

213. A nucleic acid encoding the polypeptide of claim 202.

214. A nucleic acid encoding the polypeptide of claim 204.

215. A nucleic acid encoding the polypeptide of claim 205.

216. A nucleic acid encoding the polypeptide of claim 206.

217. A nucleic acid encoding the polypeptide of claim 207.

218. A nucleic acid encoding the polypeptide of claim 208.

219. A nucleic acid encoding the polypeptide of claim 209.