WO1998038209A2

WO1998038209A2 - Secreted proteins and polynucleotides encoding them

Info

Publication number: WO1998038209A2
Application number: PCT/US1998/003697
Authority: WO
Inventors: Kenneth Jacobs; John M. Mccoy; Edward R. Lavallie; Lisa A. Racie; David Merberg; Maurice Treacy; Vikki Spaulding; Michael J. Agostino
Original assignee: Genetics Institute, Inc.
Priority date: 1997-02-26
Filing date: 1998-02-25
Publication date: 1998-09-03
Also published as: AU6439598A; EP0968287A2; WO1998038209A3; JP2002508655A; CA2281015A1

Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM

This application is a continuation-in-part of Ser. No. 60/XXX,XXX (converted to a provisional application from non-provisional application 08/805,819), filed February 26, 1997, which is incorporated by reference herein.

FIELD OF THE INVENTION The present invention provides novel polynucleotides and proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins.

BACKGROUND OF THE INVENTION Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques clone novel polynucleotides "directly" in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader sequence motif, as well as various PCR-based or low stringency hybridization cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity by virtue of their secreted nature in the case of leader sequence cloning, or by virtue of the cell or tissue source in the case of PCR-based techniques. It is to these proteins and the polynucleotides encoding them that the present invention is directed. SUMMARY OF THE INVENTION In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

^" (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 73 to nucleotide 954;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 208 to nucleotide 954; (d) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:l from nucleotide 1 to nucleotide 630;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BD380_1 deposited under accession number ATCC 98337; (f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BD380_1 deposited under accession number

ATCC 98337; (h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337;

(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity, the fragment comprising the amino acid sequence from amino acid 142 to amino acid 151 of SEQ ID NO:2;

(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above; (1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:l from nucleotide 73 to nucleotide 954; the nucleotide sequence of SEQ-ID NO:l from nucleotide 208 to nucleotide 954; the nucleotide sequence of SEQ ID NO:l from nucleotide 1 to -nucleotide 630; the nucleotide sequence of the full-length protein coding sequence of clone BD380_1 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone BD380_1 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 186.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:l.

In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:2;

(b) the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 186; (c) fragments of the amino acid sequence of SEQ ID NO:2 comprising the amino acid sequence from amino acid 142 to amino acid 151 of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:2 or the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 186.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:3;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 45 to nucleotide 281; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 61 to nucleotide 419;

(d) a polynucleotide comprising the nucleotide sequence of the full- length "protein coding sequence of clone BQ115_2 deposited under accession number ATCC 98337;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BQ115_2 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BQ115_2 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BQ115_2 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 34 to amino acid 43 of SEQ ID NO:4;

(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;

(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and

(1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:3 from nucleotide 45 to nucleotide 281; the nucleotide sequence of SEQ ID NO:3 from nucleotide 61 to nucleotide 419; the nucleotide sequence of the full-length protein coding sequence of clone BQ115_2 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone BQ115_2 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BQ115_2 deposited under accession number ATCC 98337. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4 from amino acid 7 to amino acid 79. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:3.

(a) the amino acid sequence of SEQ ID NO:4;

(b) the amino acid sequence of SEQ ID NO:4 from amino acid 7 to amino acid 79;

(c) fragments of the amino acid sequence of SEQ ID NO:4 comprising the amino acid sequence from amino acid 34 to amino acid 43 of SEQ ID NO:4; and

(d) the amino acid sequence encoded by the cDNA insert of clone BQ115J2 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:4 or the amino acid sequence of SEQ ID NO:4 from amino acid 7 to amino acid 79.

In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:5 from nucleotide 158 to nucleotide 985;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 497 to nucleotide 985;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 319 to nucleotide 923;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CC198_1 deposited under accession number ATCC 98337;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CC198_1 deposited under accession number ATCC 98337; (h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CC198_1 deposited under accession number.ATCC 98337; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6; (j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity, the fragment comprising the amino acid sequence from amino acid 133 to amino acid 142 of SEQ ID NO:6;

(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

(1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:5 from nucleotide 158 to nucleotide 985; the nucleotide sequence of SEQ ID NO:5 from nucleotide 497 to nucleotide 985; the nucleotide sequence of SEQ ID NO:5 from nucleotide 319 to nucleotide 923; the nucleotide sequence of the full-length protein coding sequence of clone CC198_1 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone CC198_1 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6 from amino acid 61 to amino acid

256.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:5.

(a) the amino acid sequence of SEQ ID NO:6;

(b) the amino acid sequence of SEQ ID NO:6 from amino acid 61 to amino acid 256; (c) fragments of the amino acid sequence of SEQ ID NO:6 comprising the amino acid sequence from amino acid 133 to amino acid 142 of-SEQ ID NO:6; and

^" -(d) the amino acid sequence encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:6 or the amino acid sequence of SEQ ID NO:6 from amino acid 61 to amino acid 256.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 21 to nucleotide 674; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:7 from nucleotide 1164 to nucleotide 1465;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337; (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 having biological activity, the fragment comprising the amino acid sequence from amino acid 104 to amino acid 113 of SEQ ID NO:8;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above; (k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and

(1) a polynucleotide capable of hybridizing under stringent conditions to any ^"one of the polynucleotides specified in (a)-(i). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:7 from nucleotide 21 to nucleotide 674; the nucleotide sequence of SEQ ID NO:7 from nucleotide 1164 to nucleotide 1465; the nucleotide sequence of the full-length protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:7. In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:8;

(b) fragments of the amino acid sequence of SEQ ID NO:8 comprising the amino acid sequence from amino acid 104 to amino acid 113 of SEQ ID NO:8; and

(c) the amino acid sequence encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:8.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:9 from nucleotide 951 to nucleotide 1037;

(c) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CS319_1 deposited under accession number ATCC 98337; (d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337;

(e) a polynucleotide comprising the nucleotide sequence of the mature protein -coding sequence of clone CS319_1 deposited under accession number ATCC 98337;

(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 9 to amino acid 18 of SEQ ID NO:10;

(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;

(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ; and

(k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:9 from nucleotide 951 to nucleotide 1037; the nucleotide sequence of the full-length protein coding sequence of clone CS319_1 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone CS319_1 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:9 or SEQ ID NO:ll.

(a) the amino acid sequence of SEQ ID NO: 10;

(b) fragments of the amino acid sequence of SEQ ID NO:10 comprising the amino acid sequence from amino acid 9 to amino acid 18 of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:10. In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:12;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:12 from nucleotide 79 to nucleotide 1134;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:12 from nucleotide 692 to nucleotide 1008;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DL504_3 deposited under accession number ATCC 98337;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DL504_3 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DL504_3 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DL504_3 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:13; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:13 having biological activity, the fragment comprising the amino acid sequence from amino acid 171 to amino acid 180 of SEQ ID NO:13;

(1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:12 from nucleotide 79 to nucleotide 1134; the nucleotide sequence of SEQ ID NO:12 from nucleotide 692 to nucleotide 1008; the nucleotide sequence of the full-length protein coding sequence of clone DL504_3 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone DL504_3 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DL504_3 deposited under accession number ATCC 98337.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:12.

(a) the amino acid sequence of SEQ ID NO:13; (b) fragments of the amino acid sequence of SEQ ID NO:13 comprising the amino acid sequence from amino acid 171 to amino acid 180 of SEQ ID NO:13; and

(c) the amino acid sequence encoded by the cDNA insert of clone DL504_3 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:13.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:14;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 14 from nucleotide 1599 to nucleotide 2543;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:14 from nucleotide 174 to nucleotide 396; (d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DN747_7 deposited under accession number ATCC 98337;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DN747_7 deposited under accession number ATCC 98337; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DN747_7 deposited under accession number ATCC 98337;

' -(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN747_^7 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:15;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:15 having biological activity, the fragment comprising the amino acid sequence from amino acid 152 to amino acid 161 of

SEQ ID NO:15;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

(1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:14 from nucleotide 1599 to nucleotide 2543; the nucleotide sequence of SEQ ID NO:14 from nucleotide 174 to nucleotide 396; the nucleotide sequence of the full-length protein coding sequence of clone DN747_7 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone DN747_7 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DN747_7 deposited under accession number ATCC 98337.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:14.

(a) the amino acid sequence of SEQ ID NO:15;

(b) fragments of the amino acid sequence of SEQ ID NO:15 comprising the amino acid sequence from amino acid 152 to amino acid 161 of SEQ ID NO:15; and (c) the amino acid sequence encoded by the cDNA insert of clone DN747_7 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:15. In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:16;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:16 from nucleotide 4 to nucleotide 1224;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 16 from nucleotide 1 to nucleotide 336;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DU123_1 deposited under accession number ATCC 98337;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DU123_1 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DU123_1 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DU123_1 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:17; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:17 having biological activity, the fragment comprising the amino acid sequence from amino acid 198 to amino acid 207 of SEQ ID NO:17;

(1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i). Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:16 from nucleotide 4 to nucleotide 1224; the nucleotide sequence of SEQ ID NO:16 from nucleotide 1 to nucleotide 336; the nucleotide sequence of the full-length protein coding sequence of clone DU123_1 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone DU123_1 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DU123_1 deposited under accession number ATCC 98337. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:17 from amino acid 1 to amino acid 111.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO: 16.

(a) the amino acid sequence of SEQ ID NO: 17;

(b) the amino acid sequence of SEQ ID NO:17 from amino acid 1 to amino acid 111; (c) fragments of the amino acid sequence of SEQ ID NO:17 comprising the amino acid sequence from amino acid 198 to amino acid 207 of SEQ ID NO:17; and

(d) the amino acid sequence encoded by the cDNA insert of clone DU123_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:17 or the amino acid sequence of SEQ ID NO:17 from amino acid 1 to amino acid 111.

NO:18;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:18 from nucleotide 233 to nucleotide 370; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18 from nucleotide 293 to nucleotide 370;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18 from nucleotide 1 to nucleotide 361; (e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337; (g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:19;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:19 having biological activity, the fragment comprising the amino acid sequence from amino acid 18 to amino acid 27 of SEQ ID NO:19;

(1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:18 from nucleotide 233 to nucleotide 370; the nucleotide sequence of SEQ ID NO:18 from nucleotide 293 to nucleotide 370; the nucleotide sequence of SEQ ID NO:18 from nucleotide 1 to nucleotide 361; the nucleotide sequence of the full-length protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337; or the nucleotide sequence of the mature protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:19 from amino acid 1 to amino acid 43. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ

ID NO:18.

In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of SEQ ID NO:19;

(b) the amino acid sequence of SEQ ID NO:19 from amino acid 1 to amino acid 43;

(c) fragments of the amino acid sequence of SEQ ID NO:19 comprising the amino acid sequence from amino acid 18 to amino acid 27 of SEQ ID NO: 19; and

(d) the amino acid sequence encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:19 or the amino acid sequence of SEQ ID NO:19 from amino acid 1 to amino acid 43.

In certain preferred embodiments, the polynucleotide is operably linked to an expression control sequence. The invention also provides a host cell, including bacterial, yeast, insect and mammalian cells, transformed with such polynucleotide compositions. Also provided by the present invention are organisms that have enhanced, reduced, or modified expression of the gene(s) corresponding to the polynucleotide sequences disclosed herein.

Processes are also provided for producing a protein, which comprise:

(a) growing a culture of the host cell transformed with such polynucleotide compositions in a suitable culture medium; and (b) purifying the protein from the culture.

The protein produced according to such methods is also provided by the present invention. Preferred embodiments include those in which the protein produced by such process is a mature form of the protein. Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier. Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention.

Method-s are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable carrier.

BRIEF DESCRIPTION OF THE DRAWINGS Figures 1 A and IB are schematic representations of the pED6 and pNOTs vectors, respectively, used for deposit of clones disclosed herein.

DETAILED DESCRIPTION ISOLATED PROTEINS AND POLYNUCLEOTIDES Nucleotide and amino acid sequences, as presently determined, are reported below for each clone and protein disclosed in the present application. The nucleotide sequence of each clone can readily be determined by sequencing of the deposited clone in accordance with known methods. The predicted amino acid sequence (both full-length and mature) can then be determined from such nucleotide sequence. The amino acid sequence of the protein encoded by a particular clone can also be determined by expression of the clone in a suitable host cell, collecting the protein and determining its sequence. For each disclosed protein applicants have identified what they have determined to be the reading frame best identifiable with sequence information available at the time of filing. As used herein a "secreted" protein is one which, when expressed in a suitable host cell, is transported across or through a membrane, including transport as a result of signal sequences in its amino acid sequence. "Secreted" proteins include without limitation proteins secreted wholly (e.g., soluble proteins) or partially (e.g. , receptors) from the cell in which they are expressed. "Secreted" proteins also include without limitation proteins which are transported across the membrane of the endoplasmic reticulum.

Clone "BD380 1"

A polynucleotide of the present invention has been identified as clone "BD380_1". BD380_1 was isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. BD380_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BD380_1 protein").

The nucleotide sequence of BD380_1 as presently determined is reported in SEQ ID NO:l. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the BD380_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:2. Amino acids 33 to 45 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 46, or are a transmembrane domain.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone BD380_1 should be approximately 1900 bp.

The nucleotide sequence disclosed herein for BD380_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BD380_1 demonstrated at least some similarity with sequences identified as AA112524 (zm28e05.rl Stratagene pancreas (#937208) Homo sapiens cDNA clone 526976 5' similar to SW:CD63_HUMAN P08962 CD63 ANTIGEN), AA113814 (zm29b04.rl Stratagene pancreas (#937208) Homo sapiens cDNA clone 527023 5'), M79068 (EST01216 Homo sapiens cDNA clone HHCPJ65), N72328 (yv31fl2.rl Homo sapiens cDNA clone), and Q61176 (Human brain Expressed Sequence Tag EST01216). The predicted amino acid sequence disclosed herein for BD380_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BD380_1 protein demonstrated at least some similarity to sequences identified as M37033 (CD53 glycoprotein [Homo sapiens]), R91446 (Human CD53 antigen), R92313 (Retinal degradation slow protein), S93788 (ocular melanoma-associated antigen, OMA81H [human,uveal]), X97227 (CD53 gene product [Mus musculus]), and Z68880 (T14G10.6 [Caenorhabditis elegans]). Based upon sequence similarity, BD380_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts four potential transmembrane domains within the

BD380_1 protein sequence, centered around amino acids 33, 73, 104, and 242 of SEQ ID NO:2, respectively. Clone "BQ115 2"

A polynucleotide of the present invention has been identified as clone "BQ115_2". BQ115_2 was isolated from a human adult colon (adenocarcinoma Caco2) cDNA library using methods-which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. BQ115_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BQ115_2 protein").

The nucleotide sequence of BQ115_2 as presently determined is reported in SEQ ID NO:3. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the BQ115_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:4.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone

BQ115_2 should be approximately 400 bp. The nucleotide sequence disclosed herein for BQ115_2 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and

FASTA search protocols. BQ115_2 demonstrated at least some similarity with sequences identified as AA130380 (zl30d03.rl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 503429 5'), N99899 (zb87el0.sl Homo sapiens cDNA clone 310602 3'), T85228 (yd33e08.rl Homo sapiens cDNA clone 110054 5'), T97822 (ye54f02.rl Homo sapiens cDNA, and U73629 (Human chromosome 11 114g8 cosmid, complete sequence). The similarity of BQ115_2 to that of the U73629 (Human chromosome 11 114g8 cosmid) sequence, the gene corresponding to BQ115_2 is likely located on human chromosome 11.

Based upon sequence similarity, BQ115_2 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts two potential transmembrane domains within the BQ115_2 protein sequence centered around amino acids 29 and 67 of SEQ ID NO:4, respectively; amino acids 55 to 67 of SEQ ID NO:4are also a possible leader /signal sequence, with the predicted mature amino acid sequence beginning at amino acid 68.

Clone "CC198 1"

A polynucleotide of the present invention has been identified as clone "CC198_1". CC198_1 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CC198_1 is_. a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CC198_1 protein"). The nucleotide sequence of CC198_1 as presently determined is reported in SEQ

ID NO:5. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CC198_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:6. Amino acids 101 to 113 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 114, or are a transmembrane domain.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CC198_1 should be approximately 1120 bp.

The nucleotide sequence disclosed herein for CC198_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CC198_1 demonstrated at least some similarity with sequences identified as AA037314 (zc52h04.rl Soares senescent fibroblasts NbHSF Homo sapiens cDNA clone 325975 5' similar to WP:T07A5.2 CE01647 YK10 HOMOLOG), AA102090 (zk87cl2.sl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 489814 3' similar to WP T07A5.2 CE01647 YK10 HOMOLOG), H94093 (yw58dl2.rl Soares placenta 8to9weeks 2NbHP8to9W Homo sapiens cDNA clone 2564395' similar to SP:YK10_YEAST

P36125 HYPOTHETICAL 32.0 KD PROTEIN IN GCN3-DAL80 INTERGENIC), N33417 (yy09a04.sl Homo sapiens cDNA clone 270702 3' similar to WP:T07A5.2 CE01647 YK10 HOMOLOG), T87106 (yd88el2.rl Homo sapiens cDNA clone 115342 5' similar to SP YK10_YEAST P36125 HYPOTHETICAL 32.0 KD PROTEIN IN GCN3-DAL80 INTERGENIC), and Z48055 (Caenorhabditis elegans cosmid T07A5). The predicted amino acid sequence disclosed herein for CC198_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CC198_1 protein demonstrated at least some similarity to sequences identified as U96638 (unc-50 related protein; URP [Rattus norvegicus]), Z48055 (T07A5.2 [Caenorhabditis elegans]), and Z95397 (unknown [Schizosaccharomyces pombe]). Based upon sequence similarity, CC198_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts five potential transmembrane domains within the CC198_1 protein sequence, centered around amino acids 110, 145, 190, 223, and 251 of SEQ ID NO:6, respectively. Clone "CT317 4"

A polynucleotide of the present invention has been identified as clone "CJ317_4". CJ317_4 was isolated from a human fetal brain cDNATibrary using methods which are selective for c^'DNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or -transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CJ317_4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CJ317_4 protein").

The nucleotide sequence of CJ317_4 as presently determined is reported in SEQ ID NO:7. What applicants presently believe to be the proper reading-frame and the predicted amino acid sequence of the CJ317_4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:8.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CJ317_4 should be approximately 2000 bp. The nucleotide sequence disclosed herein for CJ317_4 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CJ317_4 demonstrated at least some similarity with sequences identified as AA212333 (mu78d07.rl Stratagene mouse melanoma (#937312) Mus musculus cDNA clone 651661 5" similar to TR:G927407 G927407 ACTIN BINDING PROTEIN), AA251247 (zs03h05.rl NCI_CGAP_GCB1 Homo sapiens cDNA clone), D20285 (Human HL60 3'directed Mbol cDNA, HUMGS01259, clone pml854), R16712 (yf33hl2.sl Homo sapiens cDNA clone 128711 3'), R40626 (yf72gl2.sl Homo sapiens cDNA clone 28051 3'), and T20115 (Human gene signature HUMGS01259). The predicted amino acid sequence disclosed herein for CJ317_4 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CJ317_4 protein demonstrated at least some similarity to sequences identified as U18795 (Saccharomyces cerevisiae chromosome V cosmids 9669, 8334, 8199, and lambda clone 1160 [Saccharomyces cerevisiae]), U18795 (Yel057cp [Saccharomyces cerevisiae]), and X89858 (Anillin [Drosophila melanogaster]). Based upon sequence similarity, CJ317_4 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts a potential transmembrane domain within the CJ317_4 protein sequence centered around amino acid 20 of SEQ ID NO:8. Clone "CS319 1"

A polynucleotide of the present invention has been identified as clone "CS319_1".

Clones were first isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or were identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. Probes derived from these cDNAs were then used to isolate CS319_1 from a human adult testes cDNA library.

CS319_1 is a full-length clone, including the entire coding sequence of a secreted protein

(also referred to herein as "CS319_1 protein"). The nucleotide sequence of the 5' portion of CS319_1 as presently determined is reported in SEQ ID NO:9. What applicants presently believe is the proper reading frame for the coding region is indicated in SEQ ID NO:10. The predicted amino acid sequence of the CS319_1 protein corresponding to the foregoing nucleotide sequence is reported in

SEQ ID NO:10. Additional nucleotide sequence from the 3' portion of CS319_1, including the polyA tail, is reported in SEQ ID NO:ll.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CS319_1 should be approximately 1600 bp.

The nucleotide sequence disclosed herein for CS319_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CS319_1 demonstrated at least some similarity with sequences identified as AA062561 (zf68c09.rl Soares pineal gland N3HPG Homo sapiens cDNA clone 382096 5' similar to contains Ll.t2 LI repetitive element), AC000378 (*** SEQUENCING IN PROGRESS *** Human Chromosome llpl5.5 pac pDJ1173a5; HTGS phase 1, 4 unordered pieces), H04709 (ph2f06u_19/lTV Homo sapiens cDNA clone _Ph2f06u_19/1TV), L10574 (Human Chromosome 7 STS sWSS208; single read), R39402

(yh95e03.rl Homo sapiens cDNA clone 137500 5'), Z81310 (Human DNA sequence from cosmid 019A on chromosome 6 Contains HLA DNA gene and STS), Z82217 (*** SEQUENCING IN PROGRESS *** Human DNA sequence *** SEQUENCING IN PROGRESS *** from clone 78B3; HTGS phase 1), and Z95704 (Human DNA sequence from 4PTEL, Huntington's Disease Region, chromosome 4pl6.3). Based upon sequence similarity, CS319_1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of CS319_1 indicates that it may contain one or more LI repeat elements. Clone "DL504 3"

A polynucleotide of the present invention has been identified as clone "DL504_3". DL504_3 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DL504_3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DL504_3 protein").

The nucleotide sequence of DL504_3 as presently determined is reported in SEQ ID NO:12. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DL504_3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:13.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone DL504_3 should be approximately 2400 bp. The nucleotide sequence disclosed herein for DL504_3 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DL504_3 demonstrated at least some similarity with sequences identified as D81501 (Human fetal brain cDNA 5'-end GEN-168H07.1), H24009 (ym49e05.sl Homo sapiens cDNA clone 51376 3'), H50251 (yo28h02.sl Homo sapiens cDNA clone 179283 3'), R12817 (yf57bl0.rl Homo sapiens cDNA clone 262495'), U58886 (Mus musculus SH3-containing protein SH3P4 mRNA, complete cds), and X99657 (H.sapiens mRNA for protein containing SH3 domain). The predicted amino acid sequence disclosed herein for DL504_3 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted DL504_3 protein demonstrated at least some similarity to sequences identified as R71943 (Grb3-3 protein), U58886 (SH3P4 [Mus musculus]), and X99657 (SH3-containing Grb-2-like 2 [Homo sapiens]). Based upon sequence similarity, DL504_3 proteins and each similar protein or peptide may share at least some activity.

Clone "DN747 7"

A polynucleotide of the present invention has been identified as clone "DN747_7". DN747_7 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DN747_7 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DN747_7 protein").

The nucleotide sequence of DN747_7 as presently determined is reported in SEQ ID NO:14. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DN747_7 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:15.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone DN747_7 should be approximately 3450 bp. The nucleotide sequence disclosed herein for DN747_7 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DN747_7 demonstrated at least some similarity with sequences identified as AA195883 (zp92f09.rl Stratagene HeLa cell s3 937216 Homo sapiens cDNA clone 6276895' similar to WP:D2045.8 CE00608 TNF-ALPHA INDUCED PROTEIN B12), AA199716 (zq74hll.rl Stratagene hNT neuron (#937233) Homo sapiens cDNA clone 647397 5'), U55984 (Human chromosome 18ql2.1-ql2.2 clone 36 mRNA sequence), W67965 (zd39f04.sl Soares fetal heart NbHH19W Homo sapiens cDNA clone 3430393'), W68044 (zd39f04.rl Soares fetal heart NbHH19W Homo sapiens cDNA clone 343039 5'), and X79321 (R. norvegicus (Wistar) mRNA for tau microtubule-associated protein). The predicted amino acid sequence disclosed herein for DN747_7 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted DN747_7 protein demonstrated at least some similarity to sequences identified as U80842 (similar to human tumor necrosis factor-alpha-induced protein B12 (GI 179304) and several potassium channel proteins (see, U42975) [Caenorhabditis elegans]). Based upon sequence similarity, DN747_7 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts a potential transmembrane domain within the DN747_7 protein sequence around amino acid 120 of SEQ ID NO:15. The nucleotide sequence of DN747_7 indicates that it may contain one or more of the following nucleotide repeat regions: simple CCA repeat, simple AT repeat, and L1PA2.

Clone "DU123 1"

A polynucleotide of the present invention has been identified as clone "DU123_1". DU123_1 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis, of computer analysis of the amino acid sequence of the encoded protein. DU123_1 is a full-length clone, including- the entire coding sequence of a secreted protein (also referred to herein as "DU123_1 protein").

The nucleotide sequence of DU123_1 as presently determined is reported in SEQ ID NO: 16. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DU123_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:17. If a frameshift were introduced into the nucleotide sequence of SEQ ID NO:16 at position 1224, the predicted amino acid sequence for the DU123_1 protein would be extended by 96 amino acids.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone DU123_1 should be approximately 1450 bp.

The predicted amino acid sequence disclosed herein for DU123_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted DU123_1 protein demonstrated at least some similarity to sequences identified as L35240 (enigma protein [Homo sapiens]), U23769 (CLP36 [Rattus norvegicus]), and U48247 (protein kinase C-binding protein Enigma [Rattus norvegicus]). These protein sequences contain LIM domains which bind two zinc atoms and have been implicated in the recognition of tyrosine residues in tight turns. The predicted DU123_1 protein also shows some homology to human protein tyrosine phosphatase. Based upon sequence similarity, DU123_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts a potential transmembrane domain within the 20 amino terminal amino acids of the DU123_1 protein sequence of SEQ ID NO:17.

Clone "FB78 1"

A polynucleotide of the present invention has been identified as clone "FB78_1". FB78_1 was isolated from a human adult placenta cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. FB78_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "FB78_1 protein"). The nucleotide sequence of FB78_1 as presently determined is reported in SEQ ID NO:18. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the FB78_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:19. Amino acids 8 to 20 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 21, or are a transmembrane domain.

The EcoRI/NotI restriction fragment obtainable from the deposit containing clone FB78_1 should be approximately 1300 bp.

The nucleotide sequence disclosed herein for FB78_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. FB78_1 demonstrated at least some similarity with sequences identified as N50885 (yy92e05.sl Homo sapiens cDNA clone 2810243'), Q03823 (Poly GT enhancer element), T08769 (EST06661 Homo sapiens cDNA clone HIBBJ45 5' end), and U12968 (Human clone S3/1 dinucleotide repeat-rich region). Based upon sequence similarity, FB78_1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of FB78_1 indicates that it may contain a simple CA repeat sequence.

Deposit of Clones Clones BD380JL, BQ115_2, CC198_1, CJ317_4, CS319_1, DL504_3, DN747_7,

DU123_1, and FB78_1 were deposited on February 26, 1997 with the American Type Culture Collection as an original deposit under the Budapest Treaty and were given the accession number ATCC 98337, from which each clone comprising a particular polynucleotide is obtainable. All restrictions on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, except for the requirements specified in 37 C.F.R. § 1.808(b).

Each clone has been transfected into separate bacterial cells (E. coli) in this composite deposit. Each clone can be removed from the vector in which it was deposited by performing an EcoRI/NotI digestion (5' site, EcoRI; 3' site, NotI) to produce the appropriate fragment for such clone. Each clone was deposited in either the pED6 or pNOTs vector depicted in Fig. 1. The pED6dpc2 vector ("pED6") was derived from pED6dpcl by insertion of a new polylinker to facilitate cDNA cloning (Kaufman et al., 1991, Nucleic Acids Res. 19: 4485-4490); the pNOTs vector was derived from pMT2 (Kaufman et al, 1989, Mol. Cell. Biol. 9: 946-958) by deletion of the DHFR sequences, insertion of a new polylinker, and insertion of the M13 origin of replication in the Clal site. In some instances, the deposited clone can become "flipped" (i.e., in the reverse orientation) in the deposited isolate. In such instances, the cDNA insert can still be isolated by digestion with EcoRI and NotI. However, NotI will then produce the 5' site and EcoRI will produce the 3' site for- placement of the cDNA in proper orientation for expression in a suitable vector. The cDNA may also be expressed from the vectors in which they were deposited.

Bacterial cells containing a particular clone can be obtained from the composite deposit as follows:

An oligonucleotide probe or probes should be designed to the sequence that is known for that particular clone. This sequence can be derived from the sequences provided herein, or from a combination of those sequences. The sequence of the oligonucleotide probe that was used to isolate each full-length clone is identified below, and should be most reliable in isolating the clone of interest.

Clone Probe Sequence

BD380_1 SEQ ID NO:20

BQ115_2 SEQ ID NO:21 CC198_1 SEQ ID NO.22

CJ317_4 SEQ ID NO:23

CS319_1 SEQ ID NO:24

DL504_3 SEQ ID NO:25

DN747_7 SEQ ID NO:26 DU123_1 SEQ ID NO:27

FB78_1 SEQ ID NO:28

In the sequences listed above which include an N at position 2, that position is occupied in preferred probes/primers by a biotinylated phosphoaramidite residue rather than a nucleotide (such as , for example, that produced by use of biotin phosphoramidite (1- dimethoxytrityloxy-2-(N-biotinyl-4-aminobutyl)-propyl-3-0-(2-cyanoethyl)-(N,N- diisopropyl)-phosphoramadite) (Glen Research, cat. no. 10-1953)).

The design of the oligonucleotide probe should preferably follow these parameters: (a) It should be designed to an area of the sequence which has the fewest ambiguous bases ("N's"), if any;

(b) It should be designed to have a T_m of approx. 80 ° C (assuming 2° for each " A or T and 4 degrees for each G or C). The oligonucleotide should preferably be labeled with g-³²P ATP (specific activity 6000 Ci/mmole) and T4 polynucleotide kinase using commonly employed techniques for labeling oligonucleotides. Other labeling techniques can also be used. Unincorporated label should preferably be removed by gel filtration chromatography or other established methods. The amount of radioactivity incorporated into the probe should be quantitated by measurement in a scintillation counter. Preferably, specific activity of the resulting probe should be approximately 4e+6 dpm/pmole.

The bacterial culture containing the pool of full-length clones should preferably be thawed and 100 μl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 μg/ml. The culture should preferably be grown to saturation at 37°C, and the saturated culture should preferably be diluted in fresh L-broth. Aliquots of these dilutions should preferably be plated to determine the dilution and volume which will yield approximately 5000 distinct and well-separated colonies on solid bacteriological media containing L-broth containing ampicillin at 100 μg/ml and agar at 1.5% in a 150 mm petri dish when grown overnight at 37°C. Other known methods of obtaining distinct, well-separated colonies can also be employed.

Standard colony hybridization procedures should then be used to transfer the colonies to nitrocellulose filters and lyse, denature and bake them.

The filter is then preferably incubated at 65°C for 1 hour with gentle agitation in 6X SSC (20X stock is 175.3 g NaCl/liter, 88.2 g Na citrate /liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS, 100 μg/ml of yeast RNA, and 10 mM EDTA (approximately

10 mL per 150 mm filter). Preferably, the probe is then added to the hybridization mix at a concentration greater than or equal to le+6 dpm/mL. The filter is then preferably incubated at 65°C with gentle agitation overnight. The filter is then preferably washed in 500 mL of 2X SSC/0.5% SDS at room temperature without agitation, preferably followed by 500 mL of 2X SSC/0.1% SDS at room temperature with gentle shaking for 15 minutes.

A third wash with 0.1X SSC/0.5% SDS at 65°C for 30 minutes to 1 hour is optional. The filter is then preferably dried and subjected to autoradiography for sufficient time to visualize the positives on the X-ray film. Other known hybridization methods can also be employed. The positive colonies are picked, grown in culture, and plasmid DNA isolated using standard procedures. The clones can then be verified by restriction analysis, hybridization analysis, or DNA sequencing.

Fragments of the proteins of the present invention which are capable of exhibiting biological activity are also encompassed by the present invention. Fragments of the protein may be in linear form or they may be cyclized using known methods, for example, as described in H.U. Saragovi, et al., Bio /Technology 10, 773-778 (1992) and in R.S.

McDowell, et al., J. Amer. Chem. Soc. 114, 9245-9253 (1992), both of which are incorporated herein by reference. Such fragments may be fused to carrier molecules such as immunoglobulins for many purposes, including increasing the valency of protein binding sites. For example, fragments of the protein may be fused through "linker" sequences to the Fc portion of an immunoglobulin. For a bivalent form of the protein, such a fusion could be to the Fc portion of an IgG molecule. Other immunoglobulin isotypes may also be used to generate such fusions. For example, a protein - IgM fusion would generate a decavalent form of the protein of the invention.

The present invention also provides both full-length and mature forms of the disclosed proteins. The full-length form of the such proteins is identified in the sequence listing by translation of the nucleotide sequence of each disclosed clone. The mature form of such protein may be obtained by expression of the disclosed full-length polynucleotide (preferably those deposited with ATCC) in a suitable mammalian cell or other host cell. The sequence of the mature form of the protein may also be determinable from the amino acid sequence of the full-length form.

The present invention also provides genes corresponding to the polynucleotide sequences disclosed herein. "Corresponding genes" are the regions of the genome that are transcribed to produce the mRNAs from which cDNA polynucleotide sequences are derived and may include contiguous regions of the genome necessary for the regulated expression of such genes. Corresponding genes may therefore include but are not limited to coding sequences, 5' and 3' untranslated regions, alternatively spliced exons, introns, promoters, enhancers, and silencer or suppressor elements. The corresponding genes can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include the preparation of probes or primers from the disclosed sequence information for identification and /or amplification of genes in appropriate genomic libraries or other sources of genomic materials. An "isolated gene" is a gene that has been separated from the adjacent coding sequences, if any, present in the genome of the organism from which the gene was isolated.

Organisms that have enhanced, reduced, or modified expression of the gene(s) corresponding^" to the polynucleotide sequences disclosed herein are provided. The desired change in gene expression can be achieved through the use of antisense polynucleotides or ribozymes that bind and/or cleave the mRNA transcribed from the gene (Albert and Morris, 1994, Trends Pharmacol. Sci. 15(7): 250-254; Lavarosky et al., 1997, Biochem. Mol. Med. 62(1): 11-22; and Hampel, 1998, Prog. Nucleic Acid Res. Mol. Biol. 58: 1- 39; all of which are incorporated by reference herein). Transgenic animals that have multiple copies of the gene(s) corresponding to the polynucleotide sequences disclosed herein, preferably produced by transformation of cells with genetic constructs that are stably maintained within the transformed cells and their progeny, are provided. Transgenic animals that have modified genetic control regions that increase or reduce gene expression levels, or that change temporal or spatial patterns of gene expression, are also provided (see European Patent No. 0 649 464 Bl, incorporated by reference herein). In addition, organisms are provided in which the gene(s) corresponding to the polynucleotide sequences disclosed herein have been partially or completely inactivated, through insertion of extraneous sequences into the corresponding gene(s) or through deletion of all or part of the corresponding gene(s). Partial or complete gene inactivation can be accomplished through insertion, preferably followed by imprecise excision, of rransposable elements (Plasterk, 1992, Bioessays 14(9): 629-633; Zwaal et al, 1993, Proc. Natl. Acad. Sci. USA 90(16): 7431-7435; Clark et al., 1994, Proc. Natl. Acad. Sci. USA 91(2): 719-722; all of which are incorporated by reference herein), or through homologous recombination, preferably detected by positive /negative genetic selection strategies (Mansour et al, 1988, Nature 336: 348-352; U.S. Patent Nos. 5,464,764; 5,487,992; 5,627,059; 5,631,153; 5,614, 396;

5,616,491; and 5,679,523; all of which are incorporated by reference herein). These organisms with altered gene expression are preferably eukaryotes and more preferably are mammals. Such organisms are useful for the development of non-human models for the study of disorders involving the corresponding gene(s), and for the development of assay systems for the identification of molecules that interact with the protein product(s) of the corresponding gene(s).

Where the protein of the present invention is membrane-bound (e.g., is a receptor), the present invention also provides for soluble forms of such protein. In such forms part or all of the intracellular and transmembrane domains of the protein are deleted such that the protein is fully secreted from the cell in which it is expressed. The intracellular and transmembrane domains of proteins of the invention can be identified in accordance with known techniques for determination of such domains from sequence information.

Proteifis and protein fragments of the present invention include proteins with amino acid sequence lengths that are at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of a disclosed protein and have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with that disclosed protein, where sequence identity is determined by comparing the amino acid sequences of the proteins when aligned so as to maximize overlap and identity while minimizing sequence gaps. Also included in the present invention are proteins and protein fragments that contain a segment preferably comprising 8 or more (more preferably 20 or more, most preferably 30 or more) contiguous amino acids that shares at least 75% sequence identity (more preferably, at least 85% identity; most preferably at least 95% identity) with any such segment of any of the disclosed proteins. Species homologs of the disclosed polynucleotides and proteins are also provided by the present invention. As used herein, a "species homologue" is a protein or polynucleotide with a different species of origin from that of a given protein or polynucleotide, but with significant sequence similarity to the given protein or polynucleotide, as determined by those of skill in the art. Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from the desired species. Preferably, species homologs are those isolated from mammalian species.

The invention also encompasses allelic variants of the disclosed polynucleotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotide which also encode proteins which are identical, homologous, or related to that encoded by the polynucleotides .

The invention also includes polynucleotides with sequences complementary to those of the polynucleotides disclosed herein.

The present invention also includes polynucleotides capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency conditions are shown in the table below: highly stringent conditions are those that are at least as stringent as, for example, conditions A-F; stringent conditions are at least as stringent as, for example, conditions G-L, and reduced stringency conditions are at least as stringent as, for example, conditions M-R

* The hybrid length is that anticipated for the hybridized regιon(s) of the hybridizing polynucleotides When hybndizmg a polynucleotide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynucleotide When polynucleotides of known sequence are hybridized, the hybrid length can be determined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity

* SSPE (lxSSPE is 0 15M NaCl, lOmM NaH₂P0₄, and 1 25mM EDTA, pH 74) can be substituted for SSC (IxSSC is 0 15M NaCl and 15mM sodium citrate) in the hybridization and wash buffers, washes are performed for 15 minutes after hybridization is complete

T_B - T_R The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10°C less than the melting temperature (T_m) of the hybrid, where T_m is determined according to the following equations For hybrids less than 18 base pairs in length, T_m(°C) = 2(# of A + T bases) + 4(# of G + C bases). For hybrids between 18 and 49 base pairs in length, T_m(°C) = 81.5 + 16.6(log,₀[Na⁺]) + 0.41 (%G+C) - (600/N), where N is the number of bases in the hybrid, and [Na⁺] is the concentration of sodium ions in the hybridization buffer ([Na⁺] for IxSSC = 0.165 M). Additional examples of stringency conditions for polynucleotide hybridization are provided in Sa^"mbrook, J., E.F. Fritsch, and T. Maniatis, 1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology, 1995, F.M. Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4, incorporated herein by reference. Preferably, each such hybridizing polynucleotide has a length that is at least

25%(more preferably at least 50%, and most preferably at least 75%) of the length of the polynucleotide of the present invention to which it hybridizes, and has at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with the polynucleotide of the present invention to which it hybridizes, where sequence identity is determined by comparing the sequences of the hybridizing polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps.

The isolated polynucleotide of the invention may be operably linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al, Nucleic Acids Res. 19, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R. Kaufman, Methods in Enzymology 185, 537-566 (1990). As defined herein "operably linked" means that the isolated polynucleotide of the invention and an expression control sequence are situated within a vector or cell in such a way that the protein is expressed by a host cell which has been transformed (transfected) with the ligated polynucleotide/expression control sequence.

A number of types of cells may act as suitable host cells for expression of the protein. Mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, cell strains derived from in vitro culture of primary tissue, primary explants, HeLa cells, mouse L cells, BHK, HL-60, U937, HaK or Jurkat cells. Alternatively, it may be possible to produce the protein in lower eukaryotes such as yeast or in prokaryotes such as bacteria. Potentially suitable yeast strains include Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces strains, Candida, or any yeast strain capable of expressing heterologous proteins. Potentially suitable bacterial strains include Escherichia coli, Bacillus subtilis, Salmonella typhimiirium, or any bacterial strain capable of expressing heterologous proteins. If the protein is made in yeast or bacteria, it may be necessary to modify the protein produced therein, for example by phosphorylation or glycosylation of the appropriate sites, in order to obtain the functional protein. Such covalent attachments may be accomplished using known chemical or enzymatic methods.

The protein may also be produced by operably linking the isolated polynucleotide of the invention to suitable control sequences in one or more insect expression vectors, and employing an insect expression system. Materials and methods for baculovirus/insect cell expression systems are commercially available in kit form from, e.g., Invitrogen, San Diego, California, U.S.A. (the MaxBac® kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (1987), incorporated herein by reference. As used herein, an insect cell capable of expressing a polynucleotide of the present invention is "transformed." The protein of the invention may be prepared by culturing transformed host cells under culture conditions suitable to express the recombinant protein. The resulting expressed protein may then be purified from such culture (i.e., from culture medium or cell extracts) using known purification processes, such as gel filtration and ion exchange chromatography. The purification of the protein may also include an affinity column containing agents which will bind to the protein; one or more column steps over such affinity resins as concanavalin A-agarose, heparin-toyopearl® or Cibacrom blue 3GA Sepharose®; one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography. Alternatively, the protein of the invention may also be expressed in a form which will facilitate purification. For example, it may be expressed as a fusion protein, such as those of maltose binding protein (MBP), glutathione-S-transferase (GST) or thioredoxin (TRX). Kits for expression and purification of such fusion proteins are commercially available from New England BioLab (Beverly, MA), Pharmacia (Piscataway, NJ) and InVitrogen, respectively. The protein can also be tagged with an epitope and subsequently purified by using a specific antibody directed to such epitope. One such epitope ("Flag") is commercially available from Kodak (New Haven, CT).

Finally, -one or more reverse-phase high performance liquid chromatography (RP- HPLC) steps employing hydrophobic RP-HPLC media, e.g., silica gel having pendant methyl or other aliphatic groups, can be employed to further purify the protein. Some or all of the foregoing purification steps, in various combinations, can also be employed to provide a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in accordance with the present invention as an "isolated protein."

The protein of the invention may also be expressed as a product of transgenic animals, e.g., as a component of the milk of transgenic cows, goats, pigs, or sheep which are characterized by somatic or germ cells containing a nucleotide sequence encoding the protein. The protein may also be produced by known conventional chemical synthesis.

Methods for constructing the proteins of the present invention by synthetic means are known to those skilled in the art. The synthetically-constructed protein sequences, by virtue of sharing primary, secondary or tertiary structural and /or conformational characteristics with proteins may possess biological properties in common therewith, including protein activity. Thus, they may be employed as biologically active or immunological substitutes for natural, purified proteins in screening of therapeutic compounds and in immunological processes for the development of antibodies.

The proteins provided herein also include proteins characterized by amino acid sequences similar to those of purified proteins but into which modification are naturally provided or deliberately engineered. For example, modifications in the peptide or DNA sequences can be made by those skilled in the art using known techniques. Modifications of interest in the protein sequences may include the alteration, substitution, replacement, insertion or deletion of a selected amino acid residue in the coding sequence. For example, one or more of the cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Techniques for such alteration, substitution, replacement, insertion or deletion are well known to those skilled in the art (see, e.g., U.S. Patent No.4,518,584). Preferably, such alteration, substitution, replacement, insertion or deletion retains the desired activity of the protein. Other fragments and derivatives of the sequences of proteins which would be expected to retain protein activity in whole or in part and may thus be useful- for screening or other immunological methodologies may also be easily made by those skilled in the art given the disclosures herein. Such modifications are believed to be encompassed by the present invention.

USES AND BIOLOGICAL ACTIVITY

The polynucleotides and proteins of the present invention are expected to exhibit one or more of the uses or biological activities (including those associated with assays cited herein) identified below. Uses or activities described for proteins of the present invention may be provided by administration or use of such proteins or by administration or use of polynucleotides encoding such proteins (such as, for example, in gene therapies or vectors suitable for introduction of DNA).

Research Uses and Utilities

The polynucleotides provided by the present invention can be used by the research community for various purposes. The polynucleotides can be used to express recombinant protein for analysis, characterization or therapeutic use; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in disease states); as molecular weight markers on Southern gels; as chromosome markers or tags (when labeled) to identify chromosomes or to map related gene positions; to compare with endogenous DNA sequences in patients to identify potential genetic disorders; as probes to hybridize and thus discover novel, related DNA sequences; as a source of information to derive PCR primers for genetic fingerprinting; as a probe to "subtract-out" known sequences in the process of discovering other novel polynucleotides; for selecting and making oligomers for attachment to a "gene chip" or other support, including for examination of expression patterns; to raise anti-protein antibodies using DNA immunization techniques; and as an antigen to raise anti-DNA antibodies or elicit another immune response. Where the polynucleotide encodes a protein which binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the polynucleotide can also be used in interaction trap assays (such as, for example, that described in Gyuris et al, Cell 75:791-803 (1993)) to identify polynucleotides encoding the other protein with which binding occurs or to identify inhibitors of the binding interaction.

The proteins provided by the present invention can similarly be used in assay to determine biological activity, including in a panel of multiple proteins for high- throughput screening; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its receptor) in biological fluids; as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state); and, of course, to isolate correlative receptors or ligands. Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the other protein with which binding occurs or to identify inhibitors of the binding interaction. Proteins involved in these binding interactions can also be used to screen for peptide or small molecule inhibitors or agonists of the binding interaction.

Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.

Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include without limitation "Molecular Cloning: A Laboratory Manual", 2d ed., Cold Spring Harbor Laboratory Press, Sambrook,

J., E.F. Fritsch and T. Maniatis eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning Techniques", Academic Press, Berger, S.L. and A.R. Kimmel eds., 1987.

Nutritional Uses Polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. Such uses include without limitation use as a protein or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention can be added to the medium in or on which the microorganism is cultured. Cytokine and Cell Proliferation /Differentiation Activity

A protein of the present invention may exhibit cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or inhibiting) activity or may induce production of other cytokines in certain cell populations. Many protein factors discovered to date, including all known cytokines, have exhibited activity in one or more factor dependent cell proliferation assays, and hence the assays serve as a convenient confirmation of cytokine activity. The activity of a protein of the present invention is evidenced by any one of a number of routine factor dependent cell proliferation assays for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9/11, BaF3, MC9/G, M+ (preB M+), 2E8, RB5, DAI, 123, T1165, HT2, CTLL2, TF-1, Mo7e and CMK.

The activity of a protein of the invention may, among other means, be measured by the following methods:

Assays for T-cell or thymocyte proliferation include without limitation those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley- Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Bertagnolli et al, J. Immunol. 145:1706-1712, 1990; Bertagnolli et al., Cellular Immunology 133:327-341, 1991; Bertagnolli, et al., J. Immunol. 149:3778-3783, 1992; Bowman et al, J. Immunol. 152: 1756-1761, 1994.

Assays for cytokine production and /or proliferation of spleen cells, lymph node cells or thymocytes include, without limitation, those described in: Polyclonal T cell stimulation, Kruisbeek, A.M. and Shevach, E.M. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 3.12.1-3.12.14, John Wiley and Sons, Toronto. 1994; and

Measurement of mouse and human Interferon γ, Schreiber, R.D. In Current Protocols in

Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto. 1994.

Assays for proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, K., Davis, L.S. and Lipsky, P.E. In Current

Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; deVries et al., J. Exp. Med. 173:1205-1211, 1991; Moreau et al., Nature 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2931-2938, 1983; Measurement of mouse and human interleukin 6 - Nordan, R. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto. 1991; Smith et al., Proc. Natl. Acad. Sci. U.S.A. 83:1857-1861, 1986; Measurement of human Interleukin 11 - Bennett, F., Giannotti, J., Clark, S.C. and Turner, K. J. In Current Protocols in Immunology^". J.E.e.a. Coligan eds. Vol 1 pp. 6.15.1 John Wiley and Sons, Toronto. 1991; Measurement of mouse and human Interleukin 9 - Ciarletta, A., Giannotti, J., Clark, S.C. and Turner, K.J. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.13.1, John Wiley and Sons, Toronto. 1991.

Assays for T-cell clone responses to antigens (which will identify, among others, proteins that affect APC-T cell interactions as well as direct T-cell effects by measuring proliferation and cytokine production) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Immunologic studies in Humans); Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J. Immun. 11:405-411, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988.

Immune Stimulating or Suppressing Activity A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (SOD)), e.g., in regulating (up or down) growth and proliferation of T and /or B lymphocytes, as well as effecting the cytolytic activity of NK cells and other cell populations. These immune deficiencies may be genetic or be caused by viral (e.g., HIV) as well as bacterial or fungal infections, or may result from autoimmune disorders. More specifically, infectious diseases causes by viral, bacterial, fungal or other infection may be treatable using a protein of the present invention, including infections by HIV, hepatitis viruses, herpesviruses, mycobacteria, Leishmania spp., malaria spp. and various fungal infections such as candidiasis. Of course, in this regard, a protein of the present invention may also be useful where a boost to the immune system generally may be desirable, i.e., in the treatment of cancer. Autoimmune disorders which may be treated using a protein of the present invention include, for example, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, autoimmune pulmonary inflammation, Guillain-Barre^" syndrome, autoimmune thyroiditis, insulin dependent diabetes mellitis, myasthenia gravis, graft-versus-host disease and autoimmune inflammatory eye disease. Such a protein of the present invention may also to be useful in the treatment of allergic reactions and conditions, such as asthma (particularly allergic asthma) or other respiratory problems. Other conditions, in which immune suppression is desired (including, for example, organ transplantation), may also be treatable using a protein of the present invention.

Using the proteins of the invention it may also be possible to immune responses, in a number of ways. Down regulation may be in the form of inhibiting or blocking an immune response already in progress or may involve preventing the induction of an immune response. The functions of activated T cells may be inhibited by suppressing T cell responses or by inducing specific tolerance in T cells, or both. Immunosuppression of T cell responses is generally an active, non-antigen-specific, process which requires continuous exposure of the T cells to the suppressive agent. Tolerance, which involves inducing non-responsiveness or anergy in T cells, is distinguishable from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent has ceased. Operationally, tolerance can be demonstrated by the lack of a T cell response upon reexposure to specific antigen in the absence of the tolerizing agent.

Down regulating or preventing one or more antigen functions (including without limitation B lymphocyte antigen functions (such as , for example, B7)), e.g., preventing high level lymphokine synthesis by activated T cells, will be useful in situations of tissue, skin and organ transplantation and in graft-versus-host disease (GVHD). For example, blockage of T cell function should result in reduced tissue destruction in tissue transplantation. Typically, in tissue transplants, rejection of the transplant is initiated through its recognition as foreign by T cells, followed by an immune reaction that destroys the transplant. The administration of a molecule which inhibits or blocks interaction of a B7 lymphocyte antigen with its natural ligand(s) on immune cells (such as a soluble, monomeric form of a peptide having B7-2 activity alone or in conjunction with a monomeric form of a peptide having an activity of another B lymphocyte antigen (e.g., B7- 1, B7-3) or blocking antibody), prior to transplantation can lead to the binding of the molecule to the natural ligand(s) on the immune cells without transmitting the corresponding costimulatory signal. Blocking B lymphocyte antigen function in this matter prevents cytokine synthesis by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, the lack of costimulation may also be sufficient to anergize the T cells, thereby inducing tolerance in a subject. Induction of long-term tolerance by B lymphocyte antigen-blocking reagents may avoid the necessity of repeated administration of these blocking reagents. To achieve sufficient immunosuppression or tolerance in a subject, it may also be necessary to block the function of a combination of B lymphocyte antigens. The efficacy of particular blocking reagents in preventing organ transplant rejection or GVHD can be assessed using animal models that are predictive of efficacy in humans. Examples of appropriate systems which can be used include allogeneic cardiac grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which have been used to examine the immunosuppressive effects of CTLA4Ig fusion proteins in vivo as described in Lenschow et al, Science 257:789-792 (1992) and Turka et al, Proc. Natl. Acad. Sci USA, 89:11102-11105 (1992). In addition, murine models of GVHD (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 846-847) can be used to determine the effect of blocking B lymphocyte antigen function in vivo on the development of that disease. Blocking antigen function may also be therapeutically useful for treating autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against self tissue and which promote the production of cytokines and autoantibodies involved in the pathology of the diseases. Preventing the activation of autoreactive T cells may reduce or eliminate disease symptoms. Administration of reagents which block costimulation of T cells by disrupting receptor:ligand interactions of B lymphocyte antigens can be used to inhibit T cell activation and prevent production of autoantibodies or T cell-derived cytokines which may be involved in the disease process. Additionally, blocking reagents may induce antigen-specific tolerance of autoreactive T cells which could lead to long-term relief from the disease. The efficacy of blocking reagents in preventing or alleviating autoimmune disorders can be determined using a number of well-characterized animal models of human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL /Ipr/lpr mice or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB rats, and murine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 840-856).

Upregulation of an antigen function (preferably a B lymphocyte antigen function), as a means oϊ- up regulating immune responses, may also be useful in therapy. Upregulation of immune responses may be in the form of enhancing an existing immune response or eliciting an initial immune response. For example, enhancing an immune response through stimulating B lymphocyte antigen function may be useful in cases of viral infection. In addition, systemic viral diseases such as influenza, the common cold, and encephalitis might be alleviated by the administration of stimulatory forms of B lymphocyte antigens systemically.

Alternatively, anti-viral immune responses may be enhanced in an infected patient by removing T cells from the patient, costimulating the T cells in vitro with viral antigen- pulsed APCs either expressing a peptide of the present invention or together with a stimulatory form of a soluble peptide of the present invention and reintroducing the in vitro activated T cells into the patient. Another method of enhancing anti-viral immune responses would be to isolate infected cells from a patient, transfect them with a nucleic acid encoding a protein of the present invention as described herein such that the cells express all or a portion of the protein on their surface, and reintroduce the transfected cells into the patient. The infected cells would now be capable of delivering a costimulatory signal to, and thereby activate, T cells in vivo.

In another application, up regulation or enhancement of antigen function (preferably B lymphocyte antigen function) may be useful in the induction of tumor immunity. Tumor cells (e.g., sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present invention can be administered to a subject to overcome tumor-specific tolerance in the subject. If desired, the tumor cell can be transfected to express a combination of peptides. For example, tumor cells obtained from a patient can be transfected ex vivo with an expression vector directing the expression of a peptide having B7-2-like activity alone, or in conjunction with a peptide having B7-l-like activity and /or B7-3-like activity. The transfected tumor cells are returned to the patient to result in expression of the peptides on the surface of the transfected cell. Alternatively, gene therapy techniques can be used to target a tumor cell for transfection in vivo.

The presence of the peptide of the present invention having the activity of a B lymphocyte antigen(s) on the surface of the tumor cell provides the necessary costimulation signal to T cells to induce a T cell mediated immune response against the transfected tumor cells. In addition, tumor cells which lack MHC class I or. MHC class II molecules, or which fail to reexpress sufficient amounts of MHC class I or MHC class II molecules, can- be transfected with nucleic acid encoding all or a portion of (e.g., a cytoplasmic-domain truncated portion) of an MHC class I chain protein and β₂ microglobulin protein or an MHC class II α chain protein and an MHC class II β chain protein to thereby express MHC class I or MHC class II proteins on the cell surface. Expression of the appropriate class I or class II MHC in conjunction with a peptide having the activity of a B lymphocyte antigen (e.g., B7-1, B7-2, B7-3) induces a T cell mediated immune response against the transfected tumor cell. Optionally, a gene encoding an antisense construct which blocks expression of an MHC class II associated protein, such as the invariant chain, can also be cotransfected with a DNA encoding a peptide having the activity of a B lymphocyte antigen to promote presentation of tumor associated antigens and induce tumor specific immunity. Thus, the induction of a T cell mediated immune response in a human subject may be sufficient to overcome tumor-specific tolerance in the subject.

Suitable assays for thymocyte or splenocyte cytotoxicity include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M.

Kruisbeek, D.H. Margulies, E.M. Shevach, W Stiober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1- 3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc. Natl. Acad. Sci. USA 78:2488-2492, 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J. Immunol. 135:1564-1572, 1985; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al, J. Immunol. 140:508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci. USA 78:2488-2492, 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J. Immunol. 135:1564-1572, 1985; Takai et al., J. Immunol. 137:3494-3500, 1986; Bowmanet al., J. Virology 61:1992-1998; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al., Cellular Immunology 133:327-341, 1991; Brown et al., J. Immunol. 153:3079-3092, 1994.

Assays for T-cell-dependent immunoglobulin responses and isotype switching (which will identify, among others, proteins that modulate T-cell dependent antibody responses and that affect Thl/Th2 profiles) include, without limitation, those described in: Maliszewski, J. Immunol. 144:3028-3033, 1990; and Assays for B cell function: In vitro antibody production, Mond, J.J. and Brunswick, M. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto. 1994.

Mixed lymphocyte reaction (MLR) assays (which will identify, among others, proteins that generate predominantly Thl and CTL responses) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley- Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al., J. Immunol. 149:3778-3783, 1992. Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate naive T-cells) include, without limitation, those described in: Guery et al., J. Immunol. 134:536-544, 1995; Inaba et al., Journal of Experimental Medicine 173:549-559, 1991; Macatonia et al., Journal of Immunology 154:5071-5079, 1995; Porgador et al, Journal of Experimental Medicine 182:255-260, 1995; Nair et al., Journal of Virology 67:4062-4069, 1993; Huang et al., Science 264:961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169:1255-1264, 1989; Bhardwaj et al., Journal of Clinical Investigation 94:797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172:631-640, 1990.

Assays for lymphocyte survival/apoptosis (which will identify, among others, proteins that prevent apoptosis after superantigen induction and proteins that regulate lymphocyte homeostasis) include, without limitation, those described in: Darzynkiewicz et al, Cytometry 13:795-808, 1992; Gorczyca et al., Leukemia 7:659-670, 1993; Gorczyca et al., Cancer Research 53:1945-1951, 1993; Itoh et al., Cell 66:233-243, 1991; Zacharchuk, Journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993; Gorczyca et al., International Journal of Oncology 1:639-648, 1992.

Assays for proteins that influence early steps of T-cell commitment and development include, without limitation, those described in: Antica et al., Blood 84:111-117, 1994; Fine et al., Cellular Immunology 155:111-122, 1994; Galy et al., Blood 85:2770-2778, 1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551, 1991.

Hematopoiesis Regulating Activity

A protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies. Even marginal biological activity in support of colony forming cells or of factor-dependent cell lines indicates involvement in regulating hematopoiesis, e.g. in supporting the growth and proliferation of erythroid progenitor cells alone or in combination with other cytokines, thereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation /chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent myelo-suppression; in supporting the growth and proliferation of megakaryocytes and consequently of platelets thereby allowing prevention or treatment of various platelet disorders such as thrombocytopenia, and generally for use in place of or complimentary to platelet transfusions; and /or in supporting the growth and proliferation of hematopoietic stem cells which are capable of maturing to any and all of the above- mentioned hematopoietic cells and therefore find therapeutic utility in various stem cell disorders (such as those usually treated with transplantation, including, without limitation, aplastic anemia and paroxysmal nocturnal hemoglobinuria), as well as in repopulating the stem cell compartment post irradiation/chemotherapy, either in-vivo or ex-vivo (i.e., in conjunction with bone marrow transplantation or with peripheral progenitor cell transplantation (homologous or heterologous)) as normal cells or genetically manipulated for gene therapy. The activity of a protein of the invention may, among other means, be measured by the following methods:

Suitable assays for proliferation and differentiation of various hematopoietic lines are cited above.

Assays for embryonic stem cell differentiation (which will identify, among others, proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15:141-151, 1995; Keller et al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al., Blood

81:2903-2915, 1993.

Assays for stem cell survival and differentiation (which will identify, among others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylcellulose colony forming assays, Freshney, M.G. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 265-268, Wiley-Liss, Inc., New York, NY. 1994; Hirayama et al., Proc. Natl. Acad. Sci. USA 89:5907-5911, 1992; Primitive hematopoietic colony forming cells with high proliferative potential, McNiece, I.K. and Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 23-39, Wiley-Liss, Inc., New York, NY. 1994; Neben et al., Experimental Hematology 22:353-359, 1994; Cobblestone area forming cell assay, Ploemacher, R.E. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 1-21, Wiley-Liss, Inc., New York, NY. 1994; Long term bone marrow cultures in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen, T. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, Inc., New York, NY. 1994; Long term culture initiating cell assay, Sutherland, H.J. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 139-162, Wiley-Liss, Inc., New York, NY. 1994.

Tissue Growth Activity

A protein of the present invention also may have utility in compositions used for bone, cartilage, tendon, ligament and /or nerve tissue growth or regeneration, as well as for wound healing and tissue repair and replacement, and in the treatment of burns, incisions and ulcers.

A protein of the present invention, which induces cartilage and /or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals. Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.

A protein of this invention may also be used in the treatment of periodontal disease, and in other tooth repair processes. Such agents may provide an environment to attract bone-forming cells, stimulate growth of bone-forming cells or induce differentiation of progenitors of bone-forming cells. A protein of the invention may also be useful in the treatment of osteoporosis or osteoarthritis, such as through stimulation of bone and/or cartilage repair or by blocking inflammation or processes of tissue destruction (collagenase activity, osteoclast activity, etc.) mediated by inflammatory processes.

Another category of tissue regeneration activity that may be attributable to the protein of the present invention is tendon /ligament formation. A protein of the present invention, which induces tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and other animals. Such a preparation employing a tendon/ligament-like tissue inducing protein may h^'a-ve prophylactic use in preventing damage to tendon or ligament tissue, as well as use in the improved fixation of tendon or ligament to bone or other tissues, and in repairing defects to tendon or ligament tissue. De novo tendon/ligament-like tissue formation induced by a composition of the present invention contributes to the repair of congenital, trauma induced, or other tendon or ligament defects of other origin, and is also useful in cosmetic plastic surgery for attachment or repair of tendons or ligaments. The compositions of the present invention may provide an environment to attract tendon- or ligament-forming cells, stimulate growth of tendon- or ligament-forming cells, induce differentiation of progenitors of tendon- or ligament-forming cells, or induce growth of tendon /ligament cells or progenitors ex vivo for return in vivo to effect tissue repair. The compositions of the invention may also be useful in the treatment of tendinitis, carpal tunnel syndrome and other tendon or ligament defects. The compositions may also include an appropriate matrix and /or sequestering agent as a carrier as is well known in the art.

The protein of the present invention may also be useful for proliferation of neural cells and for regeneration of nerve and brain tissue, i.e. for the treatment of central and peripheral nervous system diseases and neuropathies, as well as mechanical and traumatic disorders, which involve degeneration, death or trauma to neural cells or nerve tissue. More specifically, a protein may be used in the treatment of diseases of the peripheral nervous system, such as peripheral nerve injuries, peripheral neuropathy and localized neuropathies, and central nervous system diseases, such as Alzheimer's, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome. Further conditions which may be treated in accordance with the present invention include mechanical and traumatic disorders, such as spinal cord disorders, head trauma and cerebrovascular diseases such as stroke. Peripheral neuropathies resulting from chemotherapy or other medical therapies may also be treatable using a protein of the invention.

Proteins of the invention may also be useful to promote better or faster closure of non-healing wounds, including without limitation pressure ulcers, ulcers associated with vascular insufficiency, surgical and traumatic wounds, and the like. It is expected that a protein of the present invention may also exhibit activity for generation or regeneration of other tissues, such as organs (including, for example, pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac) and vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues. Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate. A protein of the invention may also exhibit angiogenic activity.

A protein of the present invention may also be useful for gut protection or regeneration and treatment of lung or liver fibrosis, reperfusion injury in various tissues, and conditions resulting from systemic cytokine damage.

A protein of the present invention may also be useful for promoting or inhibiting differentiation of tissues described above from precursor tissues or cells; or for inhibiting the growth of tissues described above.

Assays for tissue generation activity include, without limitation, those described in: International Patent Publication No. W095/ 16035 (bone, cartilage, tendon); International Patent Publication No. WO95/05846 (nerve, neuronal); International Patent Publication No. WO91/07491 (skin, endothelium ). Assays for wound healing activity include, without limitation, those described in:

Winter, Epidermal Wound Healing, pps. 71-112 (Maibach, HI and Rovee, DT, eds.), Year Book Medical Publishers, Inc., Chicago, as modified by Eaglstein and Mertz, J. Invest. Dermatol 71:382-84 (1978).

Activin/ Inhibin Activity

A protein of the present invention may also exhibit activin- or inhibin-related activities. Inhibins are characterized by their ability to inhibit the release of follicle stimulating hormone (FSH), while activins and are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, a protein of the present invention, alone or in heterodimers with a member of the inhibin α family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals and decrease spermatogenesis in male mammals. Administration of sufficient amounts of other inhibins can induce infertility in these mammals. Alternatively, the protein of the invention, as a homodimer or as a heterodimer with other protein subunits of the inhibin- β group, may be useful as a fertility inducing therapeutic, based upon the ability of activin molecules in stimulating FSH release from cells of the anterior pituitary. See, for example, United States Patent 4,798,885. A protein of the invention may also be useful for advancement of the onset of fertility in sexually immature mammals, so as to increase the lifetime reproductive performance of domestic animals such as cows, sheep and pigs.

Assays for activin/inhibin activity include, without limitation, those described in: Vale et al., Endocrinology 91:562-572, 1972; Ling et al., Nature 321:779-782, 1986; Vale et al., Nature 321 -.776-779, 1986; Mason et al., Nature 318:659-663, 1985; Forage et al., Proc. Natl. Acad. Sci. USA 83:3091-3095, 1986.

Chemotactic/Chemokinetic Activity

A protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells. Chemotactic and chemokinetic proteins can be used to mobilize or attract a desired cell population to a desired site of action. Chemotactic or chemokinetic proteins provide particular advantages in treatment of wounds and other trauma to tissues, as well as in treatment of localized infections. For example, attraction of lymphocytes, monocytes or neutrophils to tumors or sites of infection may result in improved immune responses against the tumor or infecting agent.

A protein or peptide has chemotactic activity for a particular cell population if it can stimulate, directly or indirectly, the directed orientation or movement of such cell population. Preferably, the protein or peptide has the ability to directly stimulate directed movement of cells. Whether a particular protein has chemotactic activity for a population of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.

Assays for chemotactic activity (which will identify proteins that induce or prevent chemotaxis) consist of assays that measure the ability of a protein to induce the migration of cells across a membrane as well as the ability of a protein to induce the adhesion of one cell population to another cell population. Suitable assays for movement and adhesion include, without limitation, those described in: Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, _. Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chemokiήes 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376, 1995; Lind et al. APMIS 103:140-146, 1995; Muller et al Eur. J. Immunol. 25: 1744-1748; Gruber et al. J. of Immunol. 152:5860-5867, 1994; Johnston et al. J. of Immunol. 153: 1762-1768, 1994.

Hemostatic and Thrombolytic Activity

A protein of the invention may also exhibit hemostatic or thrombolytic activity. As a result, such a protein is expected to be useful in treatment of various coagulation disorders (including hereditary disorders, such as hemophilias) or to enhance coagulation and other hemostatic events in treating wounds resulting from trauma, surgery or other causes. A protein of the invention may also be useful for dissolving or inhibiting formation of thromboses and for treatment and prevention of conditions resulting therefrom (such as, for example, infarction of cardiac and central nervous system vessels (e.g., stroke).

Assay for hemostatic and thrombolytic activity include, without limitation, those described in: Linet et al., J. Clin. Pharmacol. 26:131-140, 1986; Burdick et al., Thrombosis

Res.45:413-419, 1987; Humphrey et al., Fibrinolysis 5:71-79 (1991); Schaub, Prostaglandins 35:467-474, 1988.

Receptor/Ligand Activity A protein of the present invention may also demonstrate activity as receptors, receptor ligands or inhibitors or agonists of receptor/ligand interactions. Examples of such receptors and ligands include, without limitation, cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, receptors involved in cell-cell interactions and their ligands (including without limitation, cellular adhesion molecules (such as selectins, integrins and their ligands) and receptor/ligand pairs involved in antigen presentation, antigen recognition and development of cellular and humoral immune responses). Receptors and ligands are also useful for screening of potential peptide or small molecule inhibitors of the relevant receptor/ligand interaction. A protein of the present invention (including, without limitation, fragments of receptors and ligands) may themselves be useful as inhibitors of receptor/ligand interactions.

The activity of a protein of the invention may, among other means, be measured by the following methods: Suitable assays for receptor-ligand activity include without limitation those described in:Current Protocols in Immunology, Ed by J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 7.28, Measurement of Cellular Adhesion under static conditions 7.28.1-7.28.22), Takai et al., Proc. Natl. Acad. Sci. USA 84:6864-6868, 1987; Bierer et al., J. Exp. Med. 168:1145-1156, 1988; Rosenstein et al, J. Exp. Med. 169:149-160 1989; Stoltenborg et al., J. Immunol. Methods 175:59-68, 1994; Stitt et al., Cell 80:661-670, 1995.

Anti-Inflammatory Activity Proteins of the present invention may also exhibit anti-inflammatory activity. The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasation, or by stimulating or suppressing production of other factors which more directly inhibit or promote an inflammatory response. Proteins exhibiting such activities can be used to treat inflammatory conditions including chronic or acute conditions), including without limitation inflammation associated with infection (such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine-induced lung injury, inflammatory bowel disease, Crohn's disease or resulting from over production of cytokines such as TNF or IL-1. Proteins of the invention may also be useful to treat anaphylaxis and hypersensitivity to an antigenic substance or material.

Cadherin /Tumor Invasion Suppressor Activity

Cadherins are calcium-dependent adhesion molecules that appear to play major roles during development, particularly in defining specific cell types. Loss or alteration of normal cadherin expression can lead to changes in cell adhesion properties linked to tumor growth and metastasis. Cadherin malfunction is also implicated in other human diseases, such as pemphigus vulgaris and pemphigus foliaceus (auto-immune blistering skin diseases), Crohn's disease, and some developmental abnormalities.

The cadherin superfamily includes well over forty members, each with a distinct pattern of expression. All members of the superfamily have in common conserved extracellular repeats (cadherin domains), but structural differences are found in other parts of the molecule. The cadherin domains bind calcium to form their tertiary structure and thus calcium is required to mediate their adhesion. Only a few amino acids in the first cadherin domain provide the basis for homophilic adhesion; modification of this recognition site can change the specificity of a cadherin so that instead of recognizing only itself, the mutant molecule can now also bind to a different cadherin. In addition, some cadherins engage in heterophilic adhesion with other cadherins.

E-cadherin, one member of the cadherin superfamily, is expressed in epithelial cell types. Pathologically, if E-cadherin expression is lost in a tumor, the malignant cells become invasive and the cancer metastasizes. Transfection of cancer cell lines with polynucleotides expressing E-cadherin has reversed cancer-associated changes by returning altered cell shapes to normal, restoring cells' adhesiveness to each other and to their substrate, decreasing the cell growth rate, and drastically reducing anchorage- independent cell growth. Thus, reintroducing E-cadherin expression reverts carcinomas to a less advanced stage. It is likely that other cadherins have the same invasion suppressor role in carcinomas derived from other tissue types. Therefore, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be used to treat cancer. Introducing such proteins or polynucleotides into cancer cells can reduce or eliminate the cancerous changes observed in these cells by providing normal cadherin expression. Cancer cells have also been shown to express cadherins of a different tissue type than their origin, thus allowing these cells to invade and metastasize in a different tissue in the body. Proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can be substituted in these cells for the inappropriately expressed cadherins, restoring normal cell adhesive properties and reducing or eliminating the tendency of the cells to metastasize.

Additionally, proteins of the present invention with cadherin activity, and polynucleotides of the present invention encoding such proteins, can used to generate antibodies recognizing and binding to cadherins. Such antibodies can be used to block the adhesion of inappropriately expressed tumor-cell cadherins, preventing the cells from forming a tumor elsewhere. Such an anti-cadherin antibody can also be used as a marker for the grade, pathological type, and prognosis of a cancer, i.e. the more progressed the cancer, the less cadherin expression there will be, and this decrease in cadherin expression can be detecte^'d by the use of a cadherin-binding antibody. Fragments of proteins of the present invention with cadherin activity, preferably a polypeptide comprising a decapeptide of the cadherin recognition site, and polynucleotides of the present invention encoding such protein fragments, can also be used to block cadherin function by binding to cadherins and preventing them from binding in ways that produce undesirable effects. Additionally, fragments of proteins of the present invention with cadherin activity, preferably truncated soluble cadherin fragments which have been found to be stable in the circulation of cancer patients, and polynucleotides encoding such protein fragments, can be used to disturb proper cell-cell adhesion.

Assays for cadherin adhesive and invasive suppressor activity include, without limitation, those described in: Hortsch et al. J Biol Chem 270 (32): 18809-18817, 1995; Miyaki et al. Oncogene 11: 2547-2552, 1995; Ozawa et al. Cell 63: 1033-1038, 1990.

Tumor Inhibition Activity

In addition to the activities described above for immunological treatment or prevention of tumors, a protein of the invention may exhibit other anti-tumor activities. A protein may inhibit tumor growth directly or indirectly (such as, for example, via

ADCC). A protein may exhibit its tumor inhibitory activity by acting on tumor tissue or tumor precursor tissue, by inhibiting formation of tissues necessary to support tumor growth (such as, for example, by inhibiting angiogenesis), by causing production of other factors, agents or cell types which inhibit tumor growth, or by suppressing, eliminating or inhibiting factors, agents or cell types which promote tumor growth.

Other Activities

A protein of the invention may also exhibit one or more of the following additional activities or effects: inhibiting the growth, infection or function of, or killing, infectious agents, including, without limitation, bacteria, viruses, fungi and other parasites; effecting

(suppressing or enhancing) bodily characteristics, including, without limitation, height, weight, hair color, eye color, skin, fat to lean ratio or other tissue pigmentation, or organ or body part size or shape (such as, for example, breast augmentation or diminution, change in bone form or shape); effecting biorhythms or caricadic cycles or rhythms; effecting the fertility of male or female subjects; effecting the metabolism, catabolism, anabolism, processing, utilization, storage or elimination of dietary fat, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional factors or component(s); effecting behavioral characteristics, including, without limitation, appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) and violent behaviors; providing analgesic effects or other pain reducing effects; promoting differentiation and growth of embryonic stem cells in lineages other than hematopoietic lineages; hormonal or endocrine activity; in the case of enzymes, correcting deficiencies of the enzyme and treating deficiency-related diseases; treatment of hyperproliferative disorders (such as, for example, psoriasis); immunoglobulin-like activity (such as, for example, the ability to bind antigens or complement); and the ability to act as an antigen in a vaccine composition to raise an immune response against such protein or another material or entity which is cross-reactive with such protein.

ADMINISTRATION AND DOSING

A protein of the present invention (from whatever source derived, including without limitation from recombinant and non-recombinant sources) may be used in a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Such a composition may also contain (in addition to protein and a carrier) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11,

IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and /or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, or to minimize side effects. Conversely, protein of the present invention may be included in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.

A protein of the present invention may be active in multimers (e.g., heterodimers or homodimers) or complexes with itself or other proteins. As a result, pharmaceutical compositions of the invention may comprise a protein of the invention in such multimeric or complexed form.

The pharmaceutical composition of the invention may be in the form of a complex of the protein(s) of present invention along with protein or peptide antigens. The protein and /or peptide antigen will deliver a stimulatory signal to both B and T lymphocytes. B lymphocytes will respond to antigen through their surface immunoglobulin receptor. T lymphocytes will respond to antigen through the T cell receptor (TCR) following presentation of the antigen by MHC proteins. MHC and structurally related proteins including those encoded by class I and class II MHC genes on host cells will serve to present the peptide antigen(s) to T lymphocytes. The antigen components could also be supplied as purified MHC-peptide complexes alone or with co-stimulatory molecules that can directly signal T cells. Alternatively antibodies able to bind surface immunolgobulin and other molecules on B cells as well as antibodies able to bind the TCR and other molecules on T cells can be combined with the pharmaceutical composition of the invention. The pharmaceutical composition of the invention may be in the form of a liposome in which protein of the present invention is combined, in addition to other pharmaceutically acceptable carriers, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Patent No. 4,235,871; U.S. Patent No. 4,501,728; U.S. Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are incorporated herein by reference. As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, i.e., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. In practicing the method of treatment or use of the present invention, a therapeutically effective amount of protein of the present invention is administered to a mammal having a condition to be treated. Protein of the present invention may be administered in accordance with the method of the invention either alone or in combination with other therapies such as treatments employing cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, protein of the present invention may be administered either simultaneously with the cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors, or sequentially. If administered sequentially, the attending physician will decide on the appropriate sequence of administering protein of the present invention in combination with cytokine(s), lymphokine(s), other hematopoietic factor(s), thrombolytic or anti-thrombotic factors.

Administration of protein of the present invention used in the pharmaceutical composition or to practice the method of the present invention can be carried out in a variety of conventional ways, such as oral ingestion, inhalation, topical application or cutaneous, subcutaneous, intraperitoneal, parenteral or intravenous injection. Intravenous administration to the patient is preferred.

When a therapeutically effective amount of protein of the present invention is administered orally, protein of the present invention will be in the form of a tablet, capsule, powder, solution or elixir. When administered in tablet form, the pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95% protein of the present invention, and preferably from about 25 to 90% protein of the present invention. When administered in liquid form, a liquid carrier such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains from about 0.5 to 90% by weight of protein of the present invention, and preferably from about 1 to 50% protein of the present invention.

When a therapeutically effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present invention will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable protein solutions, having due regard to pH, isotonicity, stability, and the like, is within the skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection should contain, in addition to protein of the present invention, an isotonic vehicle such as Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride

Injection, Lactated Ringer's Injection, or other vehicle as known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those of skill in the art.

The amount of protein of the present invention in the pharmaceutical composition of the present invention will depend upon the nature and severity of the condition being treated, and on the nature of prior treatments which the patient has undergone. Ultimately, the attending physician will decide the amount of protein of the present invention with which to treat each individual patient. Initially, the attending physician will administer low doses of protein of the present invention and observe the patient's response. Larger doses of protein of the present invention may be administered until the optimal therapeutic effect is obtained for the patient, and at that point the dosage is not increased further. It is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.01 μg to about 100 mg (preferably about O.lng to about 10 mg, more preferably about 0.1 μg to about 1 mg) of protein of the present invention per kg body weight.

The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient. It is contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration.

Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention.

Protein of the invention may also be used to immunize animals to obtain polyclonal and monoclonal antibodies which specifically react with the protein. Such antibodies may be obtained using either the entire protein or fragments thereof as an immunogen. The peptide immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, as in R.P. Merrifield, J. Amer.Chem.Soc. 85, 2149-2154 (1963); J.L. Krstenansky, et al, FEBS Lett. 211, 10 (1987). Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein. Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved. In the case of cancerous cells or leukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein.

For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering the composition topically, systematically, or locally as an implant or device. When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage. Topical administration may be suitable for wound healing and tissue repair. Therapeutically useful agents other than a protein of the invention which may also optionally be included in the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention. Preferably for bone and /or cartilage formation, the composition would include a matrix capable of delivering the protein-containing composition to the site of bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body. Such matrices may be formed of materials presently in use for other implanted medical applications. The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions will define the appropriate formulation. Potential matrices for the compositions may be biodegradable and chemically defined calcium sulfate, tricalciumphosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydrides. Other potential materials are biodegradable and biologically well- defined, such as bone or dermal collagen. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically defined, such as sintered hydroxapatite, bioglass, aluminates, or other ceramics. Matrices may be comprised of combinations of any of the above mentioned types of material, such as polylactic acid and hydroxyapatite or collagen and tricalciumphosphate. The bioceramics may be altered in composition, such as in calcium- aluminate-phosphate and processing to alter pore size, particle size, particle shape, and biodegradability. Presently preferred is a 50:50 (mole weight) copolymer of lactic acid and glycolic acid in the form of porous particles having diameters ranging from 150 to 800 microns. In some applications, it will be useful to utilize a sequestering agent, such as carboxymethyl cellulose or autologous blood clot, to prevent the protein compositions from disassociating from the matrix. A preferred family of sequestering agents is cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl- methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vinyl alcohol). The amount of sequestering agent useful herein is 0.5-20 wt%, preferably 1-10 wt% based on total formulation weight, which represents the amount necessary to prevent desorbtion of the protein from the polymer matrix and to provide appropriate handling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the osteogenic activity of the progenitor cells.

In further compositions, proteins of the invention may be combined with other agents beneficial to the treatment of the bone and /or cartilage defect, wound, or tissue in question. These agents include various growth factors such as epidermal growth factor (EGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-α and

TGF-β), and insulin-like growth factor (IGF).

The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for such treatment with proteins of the present invention. The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of a wound, type of damaged tissue (e.g., bone), the patient's age, sex, and diet, the severity of any infection, time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and with inclusion of other proteins in the pharmaceutical composition. For example, the addition of other known growth factors, such as IGF I (insulin like growth factor I), to the final composition, may also effect the dosage. Progress can be monitored by periodic assessment of tissue /bone growth and /or repair, for example, X-rays, histomorphometric determinations and tetracycline labeling.

Polynucleotides of the present invention can also be used for gene therapy. Such polynucleotides can be introduced either in vivo or ex vivo into cells for expression in a mammalian subject. Polynucleotides of the invention may also be administered by other known methods for introduction of nucleic acid into a cell or organism (including, without limitation, in the form of viral vectors or naked DNA).

Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells. Treated cells can then be introduced in vivo for therapeutic purposes.

Patent and literature references cited herein are incorporated by reference as if fully set forth.

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT : Jacobs, Kenneth McCoy, John M. LaVallie, Edward R. Racie, Lisa A. Merberg, David Treacy, Maurice Spaulding, Vikki Agostino, Michael J.

(ii) TITLE OF INVENTION: SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM

(iii) NUMBER OF SEQUENCES: 28

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: Genetics Institute, Inc.

(B) STREET: 87 CambridgePark Drive

(C) CITY: Cambridge

(D) STATE: MA

(E) COUNTRY: U.S.A.

(F) ZIP: 02140

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible

(C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: Patentin Release #1.0, Version #1.30

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: Sprunger, Suzanne A.

(B) REGISTRATION NUMBER: 41,323

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: (617) 498-8284

(B) TELEFAX: (617) 876-5851

(2) INFORMATION FOR SEQ ID NO : 1 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1694 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNEΞS : double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 1 :

CCACGAGCGC TGGCTGAGGG ACCGAGCCGG AGAGCCCCGG AGCCCCCGTA ACCCGCGCGG 60

GGAGCGCCCA GGATGCCGCG CGGGGACTCG GAGCAGGTGC GCTACTGCGC GCGCTTCTCC 120

TACCTCTGGC TCAAGTTTTC ACTTATCATC TATTCCACCG TGTTCTGGCT GATTGGGGCC 180

CTGGTCCTGT CTGTGGGCAT CTATGCAGAG GTTGAGCGGC AGAAATATAA AACCCTTGAA 240

AGTGCCTTCC TGGCTCCAGC CATCATCCTC ATCCTCCTGG GCGTCGTCAT GTTCATGGTC 300

TCCTTCATTG GTGTGCTGGC GTCCCTCCGT GACAACCTGT ACCTTCTCCA AGCATTCATG 360

TACATCCTTG GGATCTGCCT CATCATGGAG CTCATTGGTG GCGTGGTGGC CTTGACCTTC 420

CGGAACCAGA CCATTGACTT CCTGAACGAC AACATTCGAA GAGGAATTGA GAACTACTAT 480

GATGATCTGG ACTTCAAAAA CATCATGGAC TTTGTTCAGA AAAAGTTCAA GTGCTGTGGC 540

GGGGAGGACT ACCGAGATTG GAGCAAGAAT CAGTACCACG ACTGCAGTGC CCCTGGACCC 600

CTGGCCTGTG GGGTGCCCTA CACCTGCTGC ATCAGGAACA CGACAGAAGT TGTCAACACC 660

ATGTGTGGCT ACAAAACTAT CGACAAGGAG CGTTTCAGTG TGCAGGATGT CATCTACGTG 720

CGGGGCTGCA CCAACGCCGT GATCATCTGG TTCATGGACA ACTACACCAT CATGGCGGGC 780

ATCCTCCTGG GCATCCTGCT TCCCCAGTTC CTGGGGGTGC TGCTGACGCT GCTGTACATC 840

ACCCGGGTGG AGGACATCAT CATGGAGCAC TCTGTCACTG ATGGGCTCCT GGGGCCCGGT 900

GCCAAGCCCA GCGTGGAGGC GGCAGGCACG GGATGCTGCT TGTGCTACCC CAATTAGGGC 960

CCAGCCTGCC ATGGCAGCTC CAACAAGGAC CGTCTGGGAT AGCACCTCTC AGTCAACATC 1020

GTGGGGCTGG ACAGGGCTGC GGCCCCTCTG CCCACACTCA GTACTGACCA AAGCCAGGGC 1080

TGTGTGTGCC TGTGTGTAGG TCCCACGGCC TCTGCCTCCC CAGGGAGCAG AGCCTGGGCC 1140

TCCCCTAAGA GGCTTTCCCC GAGGCAGCTC TGGAATCTGT GCCCACCTGG GGCCTGGGGA 1200

ACAAGGCCCT CCTTTCTCCA GGCCTGGGCT ACGGGGGAGG GAGAGCCTGA GGCTCTGCTC 1260

AGGGCCCATT TCATCTCTGG CAGTGCCTTG GCGGTGGTAT TCAAGGCAGT TTTGTAGCAC 1320

CTGTAATTGG GGAGAGGGAG TGTGCCCCTC GGGGCAGGAG GGAAGGGCAT CTGGGGAAGG 1380

GCAGGAGGGA AGAGCTGTCC ATGCAGCCAC GCCCATGGCC AGGTTGGCCT CTTCTCAGCC 1440 TCCCAGGTGC CTTGAGCCCT CTTGCAAGGG CGGCTGCTTC CTTGAGCCTA GTTTTTTTAC 1500

GTGATTTTTG TAACATTCAT TTTTTTGTAC AGATAACAGG AGTTTCTGAC TAATCAAAGC 1560

TGGTATTTCC CCGCATGTCT TATTCTTGCC CTTCCCCCAA CCAGTTTGTT AATCAAACAA 1620

TAAAAACATG TTTTTTTTAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1680

AAAAAAAAAA AAAA 1694 (2) INFORMATION FOR SEQ ID NO : 2 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 294 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS :

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:

Met Pro Arg Gly Asp Ser Glu Gin Val Arg Tyr Cys Ala Arg Phe Ser 1 5 10 15

Tyr Leu Trp Leu Lys Phe Ser Leu lie lie Tyr Ser Thr Val Phe Trp 20 25 30

Leu He Gly Ala Leu Val Leu Ser Val Gly He Tyr Ala Glu Val Glu 35 40 45

Arg Gin Lys Tyr Lys Thr Leu Glu Ser Ala Phe Leu Ala Pro Ala He 50 55 60

He Leu He Leu Leu Gly Val Val Met Phe Met Val Ser Phe He Gly 65 70 75 80

Val Leu Ala Ser Leu Arg Asp Asn Leu Tyr Leu Leu Gin Ala Phe Met 85 90 95

Tyr He Leu Gly He Cys Leu He Met Glu Leu He Gly Gly Val Val 100 105 110

Ala Leu Thr Phe Arg Asn Gin Thr He Asp Phe Leu Asn Asp Asn He 115 120 125

Arg Arg Gly He Glu Asn Tyr Tyr Asp Asp Leu Asp Phe Lys Asn He 130 135 140

Met Asp Phe Val Gin Lys Lys Phe Lys Cys Cys Gly Gly Glu Asp Tyr 145 150 155 160 Arg Asp Trp Ser Lys Asn Gin Tyr His Asp Cys Ser Ala Pro Gly Pro 165 170 175

Leu Ala Cys Gly Val Pro Tyr Thr Cys Cys He Arg Asn Thr Thr Glu 180 185 190

Val Val Asn Thr Met Cys Gly Tyr Lys Thr He Asp Lys Glu Arg Phe 195 200 205

Ser Val Gin Asp Val He Tyr Val Arg Gly Cys Thr Asn Ala Val He 210 215 220

He Trp Phe Met Asp Asn Tyr Thr He Met Ala Gly He Leu Leu Gly 225 230 235 240

He Leu Leu Pro Gin Phe Leu Gly Val Leu Leu Thr Leu Leu Tyr He 245 250 255

Thr Arg Val Glu Asp He He Met Glu His Ser Val Thr Asp Gly Leu 260 265 270

Leu Gly Pro Gly Ala Lys Pro Ser Val Glu Ala Ala Gly Thr Gly Cys 275 280 285

Cys Leu Cys Tyr Pro Asn 290

(2) INFORMATION FOR SEQ ID NO : 3 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 419 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 3 :

GTTCTTCCGG TGGCGGAGCG GCGGATTAGC CTTCGCGGGG CAAAATGGAG CTCGAGGCCA 60

TGAGCAGATA TACCAGCCCA GTGAACCCAG CTGTCTTCCC CCATCTGACC GTGGTGCTTT 120

TGGCCATTGG CATGTTCTTC ACCGCCTGGT TCTTCGTTTA CGAGGTCACC TCTACCAAGT 180

ACACTCGTGA TATCTATAAA GAGCTCCTCA TCTCCTTAGT GGCCTCACTC TTCATGGGCT 240

TTGGAGTCCT CTTCCTGCTG CTCTGGGTTG GCATCTACGT GTGAGCACCC AAGGGTAACA 300

ACCAGATGGC TTCACTGAAA CCTGCTTTTG TAAATTACTT TTTTTTACTG TTGCTGGAAG 360

TGTCCCACCT GCTGCTCATA ATAAATGCAG ATGTATAGCA AAAAAAAAAA AAAAAAAAA 419 (2) INFORMATION FOR SEQ ID NO : 4 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 79 amino acids

(B) TYPE: amino acid (C STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 4 :

Met Glu Leu Glu Ala Met Ser Arg Tyr Thr Ser Pro Val Asn Pro Ala 1 5 10 15

Val Phe Pro His Leu Thr Val Val Leu Leu Ala He Gly Met Phe Phe 20 25 30

Thr Ala Trp Phe Phe Val Tyr Glu Val Thr Ser Thr Lys Tyr Thr Arg 35 40 45

Asp He Tyr Lys Glu Leu Leu He Ser Leu Val Ala Ser Leu Phe Met 50 55 60

Gly Phe Gly Val Leu Phe Leu Leu Leu Trp Val Gly He Tyr Val 65 70 75

( 2 ) INFORMATION FOR SEQ ID NO : 5 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1214 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 5 :

GGGAAGCCCG CCCGGTGGCG GCTGGGGTCG GCTGCTGGGA GGAGGTGGTG GGCTGGTTCG 60

GACGTGGGTC GAGGCTGTAG CAGGACTCCA GGGTTGGGAA GAACATGGAA AGTGACCTCC 120

CTGCCAAATA ACTCAGAAGA GGAGTGTCGG TAGCCAAATG TTTCTTCAGA ATACGTGTAA 180

AAGAAATGTT TTTCTTCCAT CTAGGAAGAT GTTACCGAGT ACTTCAGTGA ATTCCTTAGT 240

GCAGGGGAAC GGAGTCTTGA ATTCCAGGGA TGCGGCAAGA CACACAGCCG GAGCGAAACG 300 CTACAAATAT CTGAGAAGGC TTTTCCGCTT TCGGCAAATG GACTTTGAAT TTGCTGCCTG 360

GCAGATGCTC TACCTGTTCA CATCCCCACA GAGAGTTTAC AGAAATTTTC ATTATCGAAA 420

ACAGACGAAG GACCAGTGGG CCAGAGATGA CCCTGCTTTC TTGGTCCTGT TAAGTATCTG 480

GCTCTGTGTG TCCACTATAG GATTTGGCTT TGTGCTGGAC ATGGGATTCT TTGAGACAAT 540

AAAGCTTCTC CTTTGGGTTG TACTCATAGA TTGTGTAGGC GTTGGTCTTC TGATAGCAAC 600

TTTAATGTGG TTCATCTCTA ACAAGTATTT AGTGAAACGA CAGAGCAGAG ACTATGATGT 660

GGAATGGGGC TATGCTTTTG ATGTGCATCT CAATGCTTTT TATCCACTCC TGGTCATTTT 720

GCATTTTATC CAGCTTTTTT TCATCAACCA TGTTATCCTG ACAGACACAT TTATTGGATA 780

TTTAGTTGGA AATACCTTAT GGTTGGTTGC AGTTGGCTAT TATATCTATG TAACTTTCCT 840

GGGATACAGT GCATTGCCAT TTTTGAAAAA TACAGTAATT CTTCTGTATC CATTTGCACC 900

TCTGATTCTG CTCTACGGGC TTTCCCTGGC ACTGGGATGG AACTTCACCC ATACTCTCTG 960

TTCTTTCTAT AAGTACAGAG TGAAATAAAA AGTGAGAAGA AGATTCAATC GTAACTGTGT 1020

CAACAGTATT GTGAAGTGAT CATTTCTTGT AAAACTTGTA AATAAACTAT CATCTTTGTA 1080

GATATCTTAA AGGTGTAAAG TTTGCAAATT TGAAGAAATA TATATTAACA CTGTGGTCAG 1140

GTACATTCCT TAAAACTAAT TAAATGTACA TTTCTATAAT AAATATTTTT TAAACTAAAA 1200

AAAAAAAAAA AAAA 1214 (2) INFORMATION FOR SEQ ID NO : 6 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 276 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 6 :

Met Phe Leu Gin Asn Thr Cys Lys Arg Asn Val Phe Leu Pro Ser Arg 1 5 10 15

Lys Met Leu Pro Ser Thr Ser Val Asn Ser Leu Val Gin Gly Asn Gly 20 25 30

Val Leu Asn Ser Arg Asp Ala Ala Arg His Thr Ala Gly Ala Lys Arg 35 40 45 Tyr Lys Tyr Leu Arg Arg Leu Phe Arg Phe Arg Gin Met Asp Phe Glu 50 55 60

Phe Ala Ala Trp Gin Met Leu Tyr Leu Phe Thr Ser Pro Gin Arg Val 65 70 75 80

Tyr Arg Asn Phe His Tyr Arg Lys Gin Thr Lys Asp Gin Trp Ala Arg 85 90 95

Asp Asp Pro Ala Phe Leu Val Leu Leu Ser He Trp Leu Cys Val Ser 100 105 110

Thr He Gly Phe Gly Phe Val Leu Asp Met Gly Phe Phe Glu Thr He 115 120 125

Lys Leu Leu Leu Trp Val Val Leu He Asp Cys Val Gly Val Gly Leu 130 135 140

Leu He Ala Thr Leu Met Trp Phe He Ser Asn Lys Tyr Leu Val Lys 145 150 155 160

Arg Gin Ser Arg Asp Tyr Asp Val Glu Trp Gly Tyr Ala Phe Asp Val 165 170 175

His Leu Asn Ala Phe Tyr Pro Leu Leu Val He Leu His Phe He Gin 180 185 190

Leu Phe Phe He Asn His Val He Leu Thr Asp Thr Phe He Gly Tyr 195 200 205

Leu Val Gly Asn Thr Leu Trp Leu Val Ala Val Gly Tyr Tyr He Tyr 210 215 220

Val Thr Phe Leu Gly Tyr Ser Ala Leu Pro Phe Leu Lys Asn Thr Val 225 230 235 240

He Leu Leu Tyr Pro Phe Ala Pro Leu He Leu Leu Tyr Gly Leu Ser 245 250 255

Leu Ala Leu Gly Trp Asn Phe Thr His Thr Leu Cys Ser Phe Tyr Lys 260 265 270

Tyr Arg Val Lys 275

(2) INFORMATION FOR SEQ ID NO: 7:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1762 base pairs

(B) TYPE: nucleic acid

( C ) STRANDEDNESS : double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 7 :

GCAACATTCA TTCTTCAGTC ATGGCCAGTC CAGGAGGTCT TAGTGCTGTG CGAACCARCA 60

ACTTCGCCCT TGTTGGATCT TACACATTAT CATTGTCTTC AGTAGGAAAT ACTAAGTTTG 120

TTCTGGACAA GATAAATTAT GATGTAAGAG AGCGAGAGCT ACTGGGCTAT TTGTTCCAGG 180

AAAAGGTCCC CTTTTTATCT TCTTTGGAAG GTCATATTTA TTTAAAAATA AAATGTCAAG 240

TGAATTCCAG TGTTGAAGAA AGAGGTTTTC TAACCATATT TGAAGATGTT AGTGGTTTTG 300

GTGCCTGGCA TCGAAGATGG TGTGTTCTTT CTGGAAACTG TATATCTTAT TGGACTTATC 360

CAGATGATGA GAAACGCAAG AATCCCATAG GAAGGATAAA TCTGGCTAAT TGTACCAGTC 420

GTCAGATAGA ACCAGCCAAC AGAGAATTTT GTGCAAGACG CAACACTTTT GAATTAATTA 480

CTGTCCGACC ACAAAGAGAA GATGACCGAG AGACTCTTGT CAGCCAATGC AGGGACACAC 540

TCTGTGTTAC CAAGAACTGG CTGTCTGCAG ATACTAAAGA AGAGCGGGAT CTCTGGATGC 600

AAAAACTCAA TCAAGTTCTT GTTGATATTC GCCTCTGGCA ACCTGATGCT TGCTACGAAC 660

CTATTGGAAA GCCTTAAACC GGGAAATTTC CATGCTATCT AGAGGTTTTT GATGTCATCT 720

TAAGAAACAC ACTTAAGAGC ATCAGATTTA CTGATTGCAT TTTATGCTTT AAGTACGAAA 780

GGGTTTGTGC CAATATTCAC TACGTATTAT GCAGTATTTA TATCTTTTGT ATGTAAAACT 840

TTAACTGATT TCTGTCATTC ATCAATGAGT AGAAGTAAAT ACATTATAGT TGATTTTGCT 900

AAATCTTAAT TTAAAAGCCT CATTTTCCTA GAAATCTAAT TATTCAGTTA TTCATGACAA 960

TATTTTTTTA AAAGTAAGAA ATTCTGAGTT GTCTTCTTGG AGCTGTAGGT CTTGAAGCAG 1020

CAACGTCTTT CAGGGGTTGG AGACAGAAAC CCATTCTCCA ATCTCAGTAG TTTTTTCGAA 1080

AGGCTGTGAT CATTTATTGA TCGTGATATG ACTTGTTACT AGGGTACTGA AAAAAATGTC 1140

TAAGGCCTTT ACAGAAACAT TTTTAGTAAT GAGGATGAGA ACTTTTTCAA ATAGCAAATA 1200

TATATTGGCT TAAAGCATGA GGCTGTCTTC AGAAAAGTGA TGTGGACATA GGAGGCAATG 1260

TGTGAGACTT GGGGGTTCAA TATTTTATAT AGAAGAGTTA ATAAGCACAT GGTTTACATT 1320

TACTCAGCTA CTATATATGC AGTGTGGTGC ACATTTTCAC AGAATTCTGG CTTCATTAAG 1380

ATCATTATTT TTGCTGCGTA GCTTACAGAC TTAGCATATT AGTTTTTTCT ACTCCTACAA 1440

GTGTAAATTG AAAAATCTTT ATATTAAAAA AAGTAAACTG TTATGAAGCT GCTATGTACT 1500

AATAATACTT TGCTTGCCAA AGTGTTTGGG TTTTGTTGTT GTTTGTTTGT TTGTTTGTTT 1560 TTGGTTCATG AACAACAGTG TCTAGAAACC CATTTTGAAA GTGGAAAATT ATTAAGTCAC 1620

CTATCACCTT TAAACGCCTT TTTTTAAAAT TATAAAATAT TGTAAAGCAG GGTSTCAACT 1680

TTTAAATACA CTTTGAACTT CTTCTCTGAA TTATTAAAGT TCCTTTATGA CCTCATTTAT 1740

AAACAYTAAA AAAAAAAAAA AA 1762 (2) INFORMATION FOR SEQ ID NO : 8 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 218 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 8 :

Met Ala Ser Pro Gly Gly Leu Ser Ala Val Arg Thr Xaa Asn Phe Ala 1 5 10 15

Leu Val Gly Ser Tyr Thr Leu Ser Leu Ser Ser Val Gly Asn Thr Lys 20 25 30

Phe Val Leu Asp Lys He Asn Tyr Asp Val Arg Glu Arg Glu Leu Leu 35 40 45

Gly Tyr Leu Phe Gin Glu Lys Val Pro Phe Leu Ser Ser Leu Glu Gly 50 55 60

His He Tyr Leu Lys He Lys Cys Gin Val Asn Ser Ser Val Glu Glu 65 70 75 80

Arg Gly Phe Leu Thr He Phe Glu Asp Val Ser Gly Phe Gly Ala Trp 85 90 95

His Arg Arg Trp Cys Val Leu Ser Gly Asn Cys He Ser Tyr Trp Thr 100 105 110

Tyr Pro Asp Asp Glu Lys Arg Lys Asn Pro He Gly Arg He Asn Leu 115 120 125

Ala Asn Cys Thr Ser Arg Gin He Glu Pro Ala Asn Arg Glu Phe Cys 130 135 140

Ala Arg Arg Asn Thr Phe Glu Leu He Thr Val Arg Pro Gin Arg Glu 145 150 155 160

Asp Asp Arg Glu Thr Leu Val Ser Gin Cys Arg Asp Thr Leu Cys Val 165 170 175 Thr Lys Asn Trp Leu Ser Ala Asp Thr Lys Glu Glu Arg Asp Leu Trp 180 185 190

Met Gin Lys Leu Asn Gin Val Leu Val Asp He Arg Leu Trp Gin Pro 195 200 205

Asp Ala Cys Tyr Glu Pro He Gly Lys Pro 210 215

(2) INFORMATION FOR SEQ ID NO : 9 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1037 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:

TCGGGAGTTG GTTGCCTTTT CACTTTATTG TTTCCTTTGC TGCACAAAAT ATTTTAAGTT 60

TAATGTAGTA TAATTTATTT TAGTCTTTTT CACCTGTGCT TTTGGTTTCA TTGGACCTTC 120

TAATGCCCTG AAGCTTTTCT CTTATGTTTT CTTCTAGGAG TTTTATGGTT TTAGGCCTTA 180

TGCATAAATC TGTGAGTTAA TGTATATATA TGTTGTAAAA TAAGACCCAT CTCCATATTT 240

GGCATGTGGA TATTCAGTTT CCCAAACACC ACTTGTTGTT GTTTTATGTT TAACTTTACT 300

TTCACCAGTT TAGTATAAAG GATACAATTC AGGAACAGCC AAGAGGAAGA GATGTAAATG 360

GCAAGGAAAG ATGGAGCGAG TAGGTAACCC TGCTAAATAG CCATGATTGA GAAAATCCCC 420

CTTCAGTTGT GCTCTCCAAG AGCAGCTTAC TGGCAAAGAG ACATGCTTAC CATTATGACT 480

TATATGACTT ATATGATGCC TCACTTTGTA ACTATCACAA AGCCAGACAC AGACTCTGCA 540

AATTCCCCTT TTTCTTCCAC AATCAAATGA ACAATTTTAT CTTCAGTGGT CAGAACAAAA 600

NACTTGTTAA ACAAATTTNG CTTAAGATTC TCTCCTTCCC ACAGGCCTTG AACTTTGATT 660

CACCCTCAGT CTGAGCTTAC ATACAACTTA TCTTTATGCC TCCTCCTAAG AATAGGCTAA 720

CTTCAGGGTG AAATATCCTC TGATCTGGGA TCTAATTTTG CCACACTCCA TACTGCCCTC 780

CAGCTTTCAT CTTTTTTCAA ATCTTTTTTG NTCCTCCCTG TAAAAGGAAC CCCTTATCTT 840

CTTAACCTTT CAAATACTTG CAGATATTTT GCTTGGTGCT TCCCATCTAT TGCAATACCC 900

CTTTAGATAA AGTCAATTCT TATCTAAAAT CAAATTCATT TTATTTGACA ATGTTTACAA 960 ACAACCCCAG GACGATAACA ATTACACTCT CAATACTGGC ATCACACCTT CACAATTACA 1020 CTAACCCCAA CCTAGGC - 1037

(2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:

Met Phe Thr Asn Asn Pro Arg Thr He Thr He Thr Leu Ser He Leu 1 5 10 15

Ala Ser His Leu His Asn Tyr Thr Asn Pro Asn Leu Gly 20 25

(2) INFORMATION FOR SEQ ID NO: 11:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 419 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

TTCAGTGATC TTTTGCAAGC TCAAGATTGG CTTCAGCTGG GAGTCATTGG GCTGATGTNT 60

NTGATAACTG GTCAAGGTTA CCCACTTAGT TTTAAATGAG AAAATATCCA GGGTCTGAAG 120

AATCAAGTAA AACCACTCAA ATTCAGTTTT TATGTTTANT AAGAGGGAAG GAAATTGTAG 180

AACACTTCCA TTTNTTACCT CAACAGGAAA AAAAAAAGTA TCTATTTACT TAAGACATGT 240

AATTCAATTN TTATTTCCAG NGTAACAAAT CATTTAGTAA ACAATTATGT NTGAACACTT 300

NTAGGAGTTA CCAAATCTCA TGACATAAAA TTTAGCATTA AACTCTGACA TTCAGATGAC 360

AAAATACAGA ACAGATTTTC CACTGTCATA AATTCTCCAA TCTGAAAAAA AAAAAAAAA 419 (2) INFORMATION FOR SEQ ID NO: 12: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2536 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12:

GGCCGCCCCG CTCGGCCCTC CAGTCCCGCT CCGCCGCCTC CCTCCCGCAC AGCAGCCGCC 60

AGCGCGGCCT CCTGCACCAT GTCGGTGGCC GGCCTCAAGA AGCAGTTCCA TAAAGCCACT 120

CAGAAAGTGA GTGAGAAGGT TGGAGGAGCT GAAGGAACCA AGCTAGATGA TGACTTCAAA 180

GAGATGGAAA GGAAAGTGGA TGTCACCAGC AGGGCTGTGA TGGAAATAAT GACTAAAACA 240

ATTGAATACC TTCAACCCAA TCCAGCTTCC AGAGCTAAGC TCAGCATGAT CAACACCATG 300

TCAAAAATCC GTGGCCAGGA GAAGGGGCCA GGCTATCCTC AGGCAGAGGC GCTGCTGGCA 360

GAGGCCATGC TCAAATTTGG AAGAGAGCTT GGAGATGATT GCAACTTTGG CCCAGCACTT 420

GGTGAGGTCG GGGAGGCCAT GCGGGAACTG TCGGAGGTCA AAGACTCTTT GGACATAGAA 480

GTGAAGCAGA ACTTCATTGA CCCTCTTCAG AATCTTCATG ACAAAGATCT TAGGGAAATT 540

CAACATCATC TAAAGAAGTT GGAGGGTCGA CGCCTGGATT TTGATTATAA GAAGAAACGA 600

CAAGGCAAGA TTCCGGATGA AGAGCTTCGT CAAGCTCTAG AGAAATTTGA TGAGTCTAAG 660

GAAATTGCTG AGTCAAGCAT GTTCAATCTC TTGGAGATGG ATATTGAACA AGTGAGCCAG 720

CTCTCTGCAC TTGTGCAAGC TCAGCTGGAG TACCACAAGC AGGCAGTCCA GATCCTGCAG 780

CAAGTCACGG TCAGACTGGA AGAAAGAATA AGACAGGCTT CATCTCAGCC TAGAAGGGAA 840

TATCAACCTA AACCACGAAT GAGCCTGGAG TTTCCAACTG GAGACAGTAC TCAGCCCAAT 900

GGGGGTCTCT CCCACACAGG CACTCCCAAA CCTTCAGGTG TCCAAATGGA TCAGCCCTGC 960

TGCCGAGCTC TGTACGACTT TGAACCTGAA AATGAAGGGG AGTTGGGATT TAAAGAGGGC 1020

GATATCATCA CACTCACTAA CCAAATTGAT GAGAACTGGT ATGAGGGGAT GCTGCATGGC 1080

CATTCAGGCT TCTTCCCCAT CAATTATGTG GAAATTCTGG TTGCCCTGCC CCATTAGGAT 1140

GTTATGCTGG CTGGCTCGCC TCCTCTTGAC CCAGATAGTT ACGGTTAACC ACTGCTTTGG 1200

CAATGCTGCT TATAACACAT CCCAAGTGCA GGCCGCAGTG GTCCACGTCA TCCAGCCCCA 1260 CCAAGTGACT TTGGTTGACT TGTGGGCTCC CACAGGAGTC ATGGTGATGG ATGATATCCT 1320

CTTAGCCTGG TGGGCGTGGC ATGTGCTTTT TAAAACATCA TCTGAGACCA GCCAGTAGTC 1380

ACAGAACTGC TGTTTACACA GTTCTCAGGA GGCTGTGGTT TCTTAGAATA TGACCATGAG 1440

CCATTTCACA GAAAAACCAT CCCACCGAAG ATATTGTCTA TCACCCCAGG GGCCATCTGA 1500

AGGTCTCTTT GCATTTCTCC ATGCAAAGAG GAGAAAGCTT TTGCTTTCAC ACTGTCCCTT 1560

CCCAAATATG TGAGTCATGG AATTGTCAAA GTAAGCCTTC CCTCACCAGC AAATTGTCTC 1620

CTGATCTGAA TGAATTTGTC TCTTAATGCA TCCATAGAAA AGTGTTAATT GTGGGTTCAA 1680

AGCATTCTCT GCAAATAGGC ATCTCAGCTC CTCACACTTA TGGCTATTTC TGACGTATAG 1740

CCAGTTTTCT TCCCTCCTTG CTATTAAAGC CAGAGCGGTA ATTCCAAATT ATTTTTCAGT 1800

AAGACAGTTA ATCAGCATTA TTGTGAGAGG GACTGAAAAG AAATTCTCCA TTATGAGGAA 1860

TTGGGAAGAA ATCTGGTATC CAAGCTTAAA TTTCTTGCTA TACAGAAACT ATGTATGTAT 1920

TTAGGCTATT TCTGAAGGGC ACAGGGAAGG GGGAACAAAT ATCTTCACTT CAGTTTTATT 1980

TGTGAATTAC ATGTTTCATG AATCCATTTG GCACAGAGAC ACAAGGAAGA AAACACTAGT 2040

AACCATCTTT CCACTAGTTC ATATACTGAG AAACAGTAAA TACCTTTCCT TTCCACTTTT 2100

ACCCTGTGTT CTTTGAACAT CATTTGTGCA GATTCTGCCC TCAATGAGGA CCAAATAAAG 2160

ATGATTTTTG TGCTTAGCAG TTTAAGGTAT ATGGCTGCAT ATGCAAAACT CTTTCCCAAT 2220

TCAGTCGCTA CTTTTACTTC TGCCCTTTCT ATCCATCGTC TTCATTTTGT GTGTACAGTG 2280

CTGTGTGTAA GCTTATCAGT GTGTTTTTTT ATTTGTATCA GTCATGAAAG TCCTGTTAGG 2340

TATCCAGAGT TCTATTTATC TAGCTGTACA GACTCTTTCA GAGGTTTAAC GTGCTGCTTC 2400

CGATGTGCCA CCTGCAGTAG TGGATCATGT GGAGTGAAAG GCAAATCTTA CTGCTTAATG 2460

TATAAACTCT CACCACAGGA AGCATCGCTG TTTCCAATAA ATATTGCTGA AGACAGAAAA 2520

AAAAAAAAAA AAAAAA 2536 (2) INFORMATION FOR SEQ ID NO: 13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 352 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:

Met Ser Val Ala Gly Leu Lys Lys G-ln Phe His Lys Ala Thr Gin Lys 1 5 10 15

Val Ser Glu Lys Val Gly Gly Ala Glu Gly Thr Lys Leu Asp Asp Asp 20 ^" 25 30

Phe Lys Glu Met Glu Arg Lys Val Asp Val Thr Ser Arg Ala Val Met 35 40 45

Glu He Met Thr Lys Thr He Glu Tyr Leu Gin Pro Asn Pro Ala Ser 50 55 60

Arg Ala Lys Leu Ser Met He Asn Thr Met Ser Lys He Arg Gly Gin 65 70 75 80

Glu Lys Gly Pro Gly Tyr Pro Gin Ala Glu Ala Leu Leu Ala Glu Ala 85 90 95

Met Leu Lys Phe Gly Arg Glu Leu Gly Asp Asp Cys Asn Phe Gly Pro 100 105 HO

Ala Leu Gly Glu Val Gly Glu Ala Met Arg Glu Leu Ser Glu Val Lys 115 120 125

Asp Ser Leu Asp He Glu Val Lys Gin Asn Phe He Asp Pro Leu Gin 130 135 140

Asn Leu His Asp Lys Asp Leu Arg Glu He Gin His His Leu Lys Lys 145 150 155 160

Leu Glu Gly Arg Arg Leu Asp Phe Asp Tyr Lys Lys Lys Arg Gin Gly 165 170 175

Lys He Pro Asp Glu Glu Leu Arg Gin Ala Leu Glu Lys Phe Asp Glu 180 185 190

Ser Lys Glu He Ala Glu Ser Ser Met Phe Asn Leu Leu Glu Met Asp 195 200. 205

He Glu Gin Val Ser Gin Leu Ser Ala Leu Val Gin Ala Gin Leu Glu 210 215 220

Tyr His Lys Gin Ala Val Gin He Leu Gin Gin Val Thr Val Arg Leu 225 230 235 240

Glu Glu Arg He Arg Gin Ala Ser Ser Gin Pro Arg Arg Glu Tyr Gin 245 250 255

Pro Lys Pro Arg Met Ser Leu Glu Phe Pro Thr Gly Asp Ser Thr Gin 260 265 270

Pro Asn Gly Gly Leu Ser His Thr Gly Thr Pro Lys Pro Ser Gly Val 275 280 285

Gin Met Asp Gin Pro Cys Cys Arg Ala Leu Tyr Asp Phe Glu Pro Glu 290 295 - - ^"300

Asn Glu* Gly Glu Leu Gly Phe Lys Glu Gly Asp He He Thr Leu Thr 305 310 315 320

Asn Gin He Asp Glu Asn Trp Tyr Glu Gly Met Leu His Gly His Ser 325 330 335

Gly Phe Phe Pro He Asn Tyr Val Glu He Leu Val Ala Leu Pro His 340 345 350

(2) INFORMATION FOR SEQ ID NO: 14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3443 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14:

ACATTGCTGC CTAAAGAAAC TCAGCAGCCT CAGGCCCAAT TCTGCCACTT CTGGTTTGGG 60

TACAGTTAAA GGCAACCCTG AGGGACTTGG CAGTARAAAT CCAGGGCCTC CCCTGGGGCT 120

GGCARCTTCG TGTGCAGCTA GAGCTTTACC TGAAAGGAAG TCTCTGGGCC CAGAACTCTC 180

CACCAAGAGC CTCCCTGCCG TTCGCTGAGT CCCAGCAATT CTCCTAAGTT GAAGGGATCT 240

GAGAAGGAGA AGGAAATGTG GGGTAGATTT GGTGGTGGTT AGAGATATGC CCCCCTCATT 300

ACTGCCAACA GTTTCGGCTG CATTTCTTCA CGCACCTCGG TTCCTCTTCC TGAAGTTCTT 360

GTGCCCTGCT CTTCAGCACC ATGGGCCTTC TTATACGGAA GGCTCTGGGA TCTCCCCCTT 420

GTGGGGCAGG CTCTTGGGGC CAGCCTAAGA TCATGGTTTA GGGTGATCAG TGCTGGCAGA 480

TAAATTGAAA AGGCACGCTG GCTTGTGATC TTAAATGAGG ACAATCCCCC CAGGGCTGGG 540

CACTCCTCCC CTCCCCTCAC TTCTCCCACC TGCAGAGCCA GTGTCCTTGG GTGGGCTAAA 600

TAGGATATAC TGTATGCCGG CTCCTTCAAG CTGCTGACTC ACTTTATCAA TAGTTCCATT 660

TAAATTGACT TCAGTGGTGA GACTGTATCC TGTTTGCTAT TGCTTGTTGT GCTATGGGGG 720

GAGGGGGGAG GAATGTGTAA GATAGTTAAC ATGGGCAAAG GGAGATCTTG GGGTGCAGCA 780 CTTAAACTGC CTCGTAACCC TTTTCATGAT TTCAACCACA TTTGCTAGAG GGAGGGAGCA 840

GCCACGGAGT TAGAGGCCCT TGGGGTTTCT CTTTTCCACT GACAGGCTTT CCCAGGCAGC 900

TGGCTAGTTC ATTCCCTCCC CAGCCAGGTG CAGGCGTAGG AATATGGACA TCTGGTTGCT 960

TTGGCCTGCT GCCCTCTTTC AGGGGTCCTA AGCCCACAAT CATGCCTCCC TAAGACCTTG 1020

GCATCCTTCC CTCTAAGCCG TTGGCACCTC TGTGCCACCT CTCACACTGG CTCCAGACAC 1080

ACAGCCTGTG CTTTTGGAGC TGAGATCACT CGCTTCACCC TCCTCATCTT TGTTCTCCAA 1140

GTAAAGCCAC GAGGTCGGGG CGAGGGCAGA GGTGATCACC TGCGTGTCCC ATCTACAGAC 1200

CTGCAGCTTC ATAAAACTTC TGATTTCTCT TCAGCTTTGA AAAGGGTTAC CCTGGGCACT 1260

GGCCTAGAGC CTCACCTCCT AATAGACTTA GCCCCATGAG TTTGCCATGT TGAGCAGGAC 1320

TATTTCTGGC ACTTGCAAGT CCCATGATTT CTTCGGTAAT TCTGAGGGTG GGGGGAGGGA 1380

CATGAAATCA TCTTAGCTTA GCTTTCTGTC TGTGAATGTC TATATAGTGT ATTGTGTGTT 1440

TTAACAAATG ATTTACACTG ACTGTTGCTG TAAAAGTGAA TTTGGAAATA AAGTTATTAC 1500

TCTGATTAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAGC GGCCGCGAGA 1560

ACCAGCGTGA GTTGGAGGAG GACTCTTTTG GGCTGGCCAT GGACGAGGAC GGTCGCAAGT 1620

TCGTCTACTT CAAGTCCCTC GGGCCCTACC ACAAGTCGCG CTCGTCGTCG TGGAGCAAGA 1680

AGCGCGCCGA GAGCAGCGAC GAGGAGAACT TGCCCCGCAT GTATGAGACG GGCACCGAGT 1740

TCTGCCCCTA CGCCAGCTTC GTCAAGTACC TGTCGAAACG CAACCCTCTC TGCAAGGCGT 1800

TCTTCCAGCG GCCCCGGGAC CACTGCAGCG AGGGCGATGT GACCTGGTAC GAGAACAAAG 1860

CCATCGGCAA GAACTTGCTA GGCACTCGGA TGCAGATGCT CTCCAAGGCG GCCAAGCTCT 1920

CCAAGACCTA CACCAACCAC TGCATCGGCG CCGTCTCCAT CGCCACGCTC AACAGCATCG 1980

CGGGCATTGG CACCAAGCTG GGCTCGCCCG CCCCGCAGGG CTGCTACGCC GAGGCTCTGA 2040

ACGGGGCGGC ACGGCACAAC TCCCACCACC CCCCCACCCA TCCCTCCCAC CACCACCGCC 2100

CCCAGCCGCC CTCGCTGGGG AACACTTACA TCCTCCCCAA AGACAGCCAG GTCGGGCCCG 2160

ACGTGAAATC CGAGGCTGCG CCCAAGCGCG CCCTGTACGA GTCTGTGTTC GGGTCGGGGG 2220

AAATCTGCGG CCCCACTTCC CCCAAAAGAC TTTGTATCCG CCCCTCGGAG CCTGTGGATG 2280

CGGTGGTGGT GGTTTCCGTG AAACACGACC CCCTGCCTCT TCTTCCAGAA GCCAATGGGC 2340

ACAGAAGCAC CAATTCTCCC ACAATAGTTT CACCTGCTAT TGTTTCCCCC ACCCAGGACA 2400

GTCGGCCCAA TATGTCAAGA CCTCTGATCA CTAGATCCCC TGCATCTCCA CTGAACAACC 2460 AAGGCATCCC TACTCCAGCA CAACTCACAA AATCCAATGC GCCTGTCCAC ATTGATGTGG 2520

GCGGCCACAT GTACACCAGC AGCSTGGCCA CCCTCACCAA ATACCCTGAA TCCAGAATCG 2580

GAAGACTTTT TGATGGTACA GAGCCCATTG TTTTGGACAG TCTCARACAG CACTATTTCA 2640

TTGACAGAGA TGGACAGATG TTCAGATATA TCTTGAATTT TCTACGAACA TCCAAACTCC 2700

TCATTCCTGA TGATTTCAAG GACTACACTT TGTTATATGA AGAGGCAAAA TATTTTCAGC 2760

TTCAGCCCAT GTTGTTGGAG ATGGAAAGAT GGAAGCAGGA CAGAGAAACT GGTCGATTTT 2820

CAAGGCCCTG TGAGTGCCTC GTCGTGCGTG TGGCCCCAGA CCTCGGAGAA AGGATCACGC 2880

TAAGCGGTGA CAAATCCTTG ATAGAAGAAG TATTTCCAGA GATCGGCGAC GTGATGTGTA 2940

ACTCTGTCAA TGCAGGCTGG AATCACGACT CGACGCACGT CATCAGGTTT CCACTAAATG 3000

GCTACTGTCA CCTCAACTCA GTCCAGGTCC TCGAGAGGTT GCAGCAAAGA GGATTTGAAA 3060

TCGTGGGCTC CTGTGGGGGA GGAGTAGACT CGTCCCAGTT CAGCGAATAC GTCCTTCGGC 3120

GGGAACTGAG GCGGACGCCC CGTGTACCCT CCGTCATCCG GATAAAGCAA GAGCCTCTGG 3180

ACTAAATGGA CATATTTCTT ATGCAAAAAG GAAAACACAC ACAACCAATA ACTCAAACAA 3240

AAAAGGGACA TTTATGTGCA GTTGGGACAG CAAACCAAGT CCTGGACGTA AAATCGAATA 3300

AAAGACACAT TTATATCCAA TAGAGACCAC ACCTGTATTC ATATGGGAAC AATTGGAATA 3360

GTGATATCCT CAAGGTGTAA AAAATATATA AATATATATA TATATGTCAA AAGGTAGGAA 3420

ATGCAAAAAA AAAAAAAAAA AAA 3443 (2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 315 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

Met Asp Glu Asp Gly Arg Lys Phe Val Tyr Phe Lys Ser Leu Gly Pro 1 5 10 15

Tyr His Lys Ser Arg Ser Ser Ser Trp Ser Lys Lys Arg Ala Glu Ser 20 25 30 Ser Asp Glu Glu Asn Leu Pro Arg Met Tyr Glu Thr Gly Thr Glu Phe 35 40 45

Cys Pro Tyr Ala Ser Phe Val Lys Tyr _Leu_ Ser Lys Arg Asn Pro Leu 50 55 60

Cys Lys Ala Phe Phe Gin Arg Pro Arg Asp His Cys Ser Glu Gly Asp 65 70 ^' 75 80

Val Thr Trp Tyr Glu Asn Lys Ala He Gly Lys Asn Leu Leu Gly Thr 85 90 95

Arg Met Gin Met Leu Ser Lys Ala Ala Lys Leu Ser Lys Thr Tyr Thr 100 105 110

Asn His Cys He Gly Ala Val Ser He Ala Thr Leu Asn Ser He Ala 115 120 125

Gly He Gly Thr Lys Leu Gly Ser Pro Ala Pro Gin Gly Cys Tyr Ala 130 135 140

Glu Ala Leu Asn Gly Ala Ala Arg His Asn Ser His His Pro Pro Thr 145 150 155 160

His Pro Ser His His His Arg Pro Gin Pro Pro Ser Leu Gly Asn Thr 165 170 175

Tyr He Leu Pro Lys Asp Ser Gin Val Gly Pro Asp Val Lys Ser Glu 180 185 190

Ala Ala Pro Lys Arg Ala Leu Tyr Glu Ser Val Phe Gly Ser Gly Glu 195 200 205

He Cys Gly Pro Thr Ser Pro Lys Arg Leu Cys He Arg Pro Ser Glu 210 215 220

Pro Val Asp Ala Val Val Val Val Ser Val Lys His Asp Pro Leu Pro 225 230 235 240

Leu Leu Pro Glu Ala Asn Gly His Arg Ser Thr Asn Ser Pro Thr He 245 250 255

Val Ser Pro Ala He Val Ser Pro Thr Gin Asp Ser Arg Pro Asn Met 260 265 270

Ser Arg Pro Leu He Thr Arg Ser Pro Ala Ser Pro Leu Asn Asn Gin 275 280 285

Gly He Pro Thr Pro Ala Gin Leu Thr Lys Ser Asn Ala Pro Val His 290 295 300

He Asp Val Gly Gly His Met Tyr Thr Ser Ser 305 310 315

(2) INFORMATION FOR SEQ ID NO: 16: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1511 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:

AACATGGGCA CAGGGGATTT TATCTGCATT TCCATGACTG GAGGGGCGCC CTGGGGGTTC 60

AGATTGCAAG GTGGCAAGGA GCAGAAGCAG CCCTTACAAG TTGCAAAGAT TCGAAATCAG 120

AGCAAAGCCT CTGGGTCTGG GCTCTGTGAG GGAGATGAAG TGGTTTCCAT CAATGGCAAC 180

CCTTGTGCAG ATCTCACCTA CCCTGAAGTC ATCAAGCTCA TGGAAAGCAT AACAGACTCT 240

CTCCAAATGC TCATCAAAAG ACCATCCAGT GGAATAAGTG AGGCTTTGAT ATCTGAAAAT 300

GAAAACAAAA ACCTCGAGCA TCTCACACAT GGGGGTTATG TGGAAAGTAC CACCCTGCAG 360

ATTCGACCGG CCACAAAGAC CCAGTGCACA GAATTCTTCC TCGCCCCTGT CAAGACTGAA 420

GTTCCCCTAG CTGAGAACCA AAGAAGTGGT CCCGACTGTG CAGGCAGCTT GAAAGAAGAA 480

ACAGGCCCGA GCTACCAAAG GGCTCCCCAA ATGCCTGACT CCCAAAGAGG ACGCGTGGCA 540

GAAGAGCTGA TCTTAAGGGA GAAGGTAGAA GCGGTACAGC CTGGGCCTGT GGTTGAGCTG 600

CAACTGTCCC TTTCACAGGA GAGACATAAG GGCGCTAGTG GCCCTTTAGT GGCTCTCCCG 660

GGAGCTGAAA AATCTAAGTC TCCTGACCCA GACCCTAACT TGTCACATGA CAGGATTGTC 720

CACATAAATT CGATCCCTAC TAATGAGAAA GCAGACCCTT TCCTGAGGTC CAGCAAGATA 780

ATCCAGATCT CCAGTGGCAG AGAGTTGAGA GTGATCCAGG AAAGTGAAGC AGGAGATGCG 840

GGACTGCCCC GGGTGGAAGT GATCCTCGAC TGCTCTGACA GGCAGAAGAC AGAAGGGTGC 900

AGGCTTCAGG CAGGAAAGGA GTGTGTGGAT TCTCCAGTGG AAGGAGGGCA GTCAGAAGCA 960

CCTCCTTCTC TGGTATCCTT TGCCGTCTCA TCAGAAGGCA CAGAGCAGGG AGAAGATCCA 1020

CGCTCGGAAA AAGATCACAG CAGACCTCAC AAGCACCGAG CGCGGCATGC ACGGCTCAGG 1080

AGGAGTGAAA GCCTGTCAGA AAAACAAGTG AAGGAAGCAA AATCTAAATG CAAAAGCATT 1140

GCCCTTCTTC TAACGGATGC TCCCAACCCC AAYTCCAAGG GGGTGTTGAT GTTTAAGAAG 1200

CGACGTCGGA GGGCCAGGAA ATAACACCCT AGTTAGCTAC GGTACTGGCG AGCTTGAGCG 1260 AGAGGCGGAC GAGGAGGAAG AAGGTGACAA GGAGGATACA TGTGAAGTAG CATTTCTTGG 1320

TGCAAGCGAA TCAGAGGTGG ATGAAGAGTT ATTGTCTGAC GTTGACGACA ACACACAAGT 1380

TGTGAACTTT GACTGGGATT CTGGACTGGT GGACATTGAA AAGAAACTGA ACAGAGGGGA 1440

CAAGATGGAG ATGTTACCAG ACACCACAGG CAAGGGAGAC ACTGACTCCA GCCAAGAAAA 1500

AAAAAAAAAA A 1511 (2) INFORMATION FOR SEQ ID NO: 17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 406 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17:

Met Gly Thr Gly Asp Phe He Cys He Ser Met Thr Gly Gly Ala Pro 1 5 10 15

Trp Gly Phe Arg Leu Gin Gly Gly Lys Glu Gin Lys Gin Pro Leu Gin 20 25 30

Val Ala Lys He Arg Asn Gin Ser Lys Ala Ser Gly Ser Gly Leu Cys 35 40 45

Glu Gly Asp Glu Val Val Ser He Asn Gly Asn Pro Cys Ala Asp Leu 50 55 60

Thr Tyr Pro Glu Val He Lys Leu Met Glu Ser He Thr Asp Ser Leu 65 70 75 80

Gin Met Leu He Lys Arg Pro Ser Ser Gly He Ser Glu Ala Leu He 85 90 95

Ser Glu Asn Glu Asn Lys Asn Leu Glu His Leu Thr His Gly Gly Tyr 100 105 110

Val Glu Ser Thr Thr Leu Gin He Arg Pro Ala Thr Lys Thr Gin Cys 115 120 125

Thr Glu Phe Phe Leu Ala Pro Val Lys Thr Glu Val Pro Leu Ala Glu 130 135 140

Asn Gin Arg Ser Gly Pro Asp Cys Ala Gly Ser Leu Lys Glu Glu Thr 145 150 155 160 Gly Pro Ser Tyr Gin Arg Ala Pro Gin Met Pro Asp Ser Gin Arg Gly 165 170 175

Arg Val Ala Glu Glu Leu He Leu Arg Glu Lys Val Glu Ala Val Gin 180 185 190

Pro Gly Pro Val Val Glu Leu Gin Leu Ser Leu Ser Gin Glu Arg His 195 200 205

Lys Gly Ala Ser Gly Pro Leu Val Ala Leu Pro Gly Ala Glu Lys Ser 210 215 220

Lys Ser Pro Asp Pro Asp Pro Asn Leu Ser His Asp Arg He Val His 225 230 235 240

He Asn Ser He Pro Thr Asn Glu Lys Ala Asp Pro Phe Leu Arg Ser 245 250 255

Ser Lys He He Gin He Ser Ser Gly Arg Glu Leu Arg Val He Gin 260 265 270

Glu Ser Glu Ala Gly Asp Ala Gly Leu Pro Arg Val Glu Val He Leu 275 280 285

Asp Cys Ser Asp Arg Gin Lys Thr Glu Gly Cys Arg Leu Gin Ala Gly 290 295 300

Lys Glu Cys Val Asp Ser Pro Val Glu Gly Gly Gin Ser Glu Ala Pro 305 310 315 320

Pro Ser Leu Val Ser Phe Ala Val Ser Ser Glu Gly Thr Glu Gin Gly 325 330 335

Glu Asp Pro Arg Ser Glu Lys Asp His Ser Arg Pro His Lys His Arg 340 345 350

Ala Arg His Ala Arg Leu Arg Arg Ser Glu Ser Leu Ser Glu Lys Gin 355 360 365

Val Lys Glu Ala Lys Ser Lys Cys Lys Ser He Ala Leu Leu Leu Thr 370 375 380

Asp Ala Pro Asn Pro Asn Ser Lys Gly Val Leu Met Phe Lys Lys Arg 385 390 395 400

Arg Arg Arg Ala Arg Lys 405

(2) INFORMATION FOR SEQ ID NO: 18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1329 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18:

CCTTCTTCCT CACTCACACA TTTTTTGTAC ATCTGGGCCC TTAGTTTTTA TTCTGTTTAT 60

TATATGTCTC TGTCTCTCTC TATTGTGTGT GTGTGTGTGT GTGTGTGTGT GTGTGTGTGT 120

GTGTGTGTGT GGTGCAGGAG TGCCACCCCC AGGGCCCTGT CAACCTCTCT TTTCTCCTCC 180

ATGGCTGTCT GCCTGCGTAT CTGTCTCTGA GAATCCTCGG GGCGGTCAGG GGATGTCAGG 240

AGGGGAAGGA GCCGCCCTCC CTATCTTGCT GCTCCTCTTG GCACTCAGGG GCACCTTCCA 300

TGGAGCCAGA CCGGGTGGAG GGGCTTCTGG GATTTGGTGT CTGCTGCTGC CAGAGCAGGA 360

ACCCCCAGTC TAGGACTTGG GCATTTTAAC AGGGAGAAAG TAGTGGCTTC CCTTTTCTCT 420

CTCTCCTCCT TTTTCCCTTT AAGCCCACAG ATTCAGGTCA TGCCAAAAGC TCTCTGGTTG 480

TAACCTGGAG ACATGTGGAG GGGAATGGCG ATGGGATTAT AGGACTCTCC CCATCTCGGG 540

CCCTGACCCT GACCCTTGCC ACCAACCCAA AGACAGCTGG TGGGTTTCCC CTTGGAGACA 600

ATCCTGCGTT TGCCTGGGCC GGCCCTGGCT GCCCTCAGCT TTCGCTGATC TGCCCGGCCT 660

GGAGCCTCCC ATCACCCCGC TTCTTGTTGG GCCTCAGGCA CTGGTTACCA GAAGGGGGTC 720

TGGGTCTGCT CAGGATCATG TTTTGTAGCA CCTCCTGTTG GAGGGGTGGA GGGATGTTCC 780

CCTGAGCCAG GCTGAGACTA GAACCCCATC TTCCCTGAGC CAGGCTGAGA CTAGAACCCC 840

ATCTTCCCCA CCACGCCACC CCTGTGGCTG CTACAGGAGC ACAGTAGTGA AGGCCTGAGC 900

TCCAGGTTTG AAAGACCCAA CTGGAGCGTG GGGCGGGCAG GCAGGGGTTA GTGAAAGGAC 960

ACTTCCAGGG TTAGGACAGA GCATTTAGCC TTCTGGAAGA ACCCCTGCCT GGGGTGGGAC 1020

TGTGCAGGCC AGAGAAGGTG GCATGGGCCT GAACCCACCT GGACTGACTT CTGCACTGAA 1080

GCCACAGATG GAGGGTAGGC TGGTGGGTGG GGGTGGTTCG TTCTCTAGCC GGGGCAGACA 1140

CCCAGCTGGC TGGGTCCTTC YTCAGCCTTG CCTCCTCCTG TCCCCAACCC TTTCCTTTCC 1200

TCCTGCTTGC GGACTGCTGG TCCCCTCTCC TTCCCTCCTT CCAGCTGTTT CTAGTTACCA 1260

CCTACCCCTG GCCGTGGACT GATCAGACCA GCATTCAAAA TAAAAGTTTG TTCCAAAAAA 1320

AAAAAAAAA 1329 (2) INFORMATION FOR SEQ ID NO: 19: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 46 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:

Met Ser Gly Gly Glu Gly Ala Ala Leu Pro He Leu Leu Leu Leu Leu 1 5 10 15

Ala Leu Arg Gly Thr Phe His Gly Ala Arg Pro Gly Gly Gly Ala Ser 20 25 30

Gly He Trp Cys Leu Leu Leu Pro Glu Gin Glu Pro Pro Val 35 40 45

[ 2 ) INFORMATION FOR SEQ ID NO: 20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: ANCCCAAGGAT GTACATGAAT GCTTGGAG 29

(2) INFORMATION FOR SEQ ID NO: 21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: GNTCACTGGGC TGGTATATCT GCTCACGT 29

(2) INFORMATION FOR SEQ ID NO: 22:

(i) SEQUENCE CHARACTERISTICS: (A.) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: CNGGATAAAAT GCAAAATGAC CAGGAGTG 29

(2) INFORMATION FOR SEQ ID NO: 23:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23: ANATGGGTTTC TAGACACTGT TGTTCATG 29

(2) INFORMATION FOR SEQ ID NO: 24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: GNGAAGCACCA AGCAAAATAT CTGCAAGT 29 (2) INFORMATION FOR SEQ ID NO: 25:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid (C-) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25: ANCCGTAACTA TCTGGGTCAA GAGGAGGC 29

(2) INFORMATION FOR SEQ ID NO: 26:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26: CNGGAATGAGG AGTTTGGATG TTCGTAGA 29

(2) INFORMATION FOR SEQ ID NO: 27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: TNGCAACTTGT AAGGGCTGCT TCTGCTCC 29

(2) INFORMATION FOR SEQ ID NO: 28: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: other nucleic acid

(A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28: CNCAGAGACAG ATACGCAGGC AGACAGCC 29

Claims

What is claimed is:

1. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l;^* -

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 208 to nucleotide 954;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 1 to nucleotide 630;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BD380_1 deposited under accession number ATCC 98337;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BD380_1 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337;

(m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).

2. The polynucleotide of claim 1 wherein said polynucleotide is operably linked to at least one expression control sequence.

3. ^" A host cell transformed with the polynucleotide of claim 2.

4. The host cell of claim 3, wherein said cell is a mammalian cell.

5. A process for producing a protein encoded by the polynucleotide of claim 2, which process comprises:

(a) growing a culture of the host cell of claim 3 in a suitable culture medium; and

(b) purifying said protein from the culture.

6. A protein produced according to the process of claim 5.

7. The protein of claim 6 comprising a mature protein.

8. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:2;

(b) the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 186;

(c) fragments of the amino acid sequence of SEQ ID NO:2 comprising the amino acid sequence from amino acid 142 to amino acid 151 of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone BD380_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

9. The protein of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2.

10. The protein of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 186.

11. A composition comprising the protein of claim 8 and a pharmaceutically acceptable carrier.

12. ^" A method for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition of claim 11.

13. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:l.

14. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 45 to nucleotide 281;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 61 to nucleotide 419;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BQ115_2 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BQ115J2 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 34 to amino acid 43 of SEQ ID NO:4;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

15. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:4;

(b) the amino acid sequence of SEQ ID NO:4 from amino acid 7 to amino acid 79;

(d) the amino acid sequence encoded by the cDNA insert of clone BQ115_2 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

16. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:3.

17. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 from nucleotide 158 to nucleotide 985;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337; (g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CC198_1 deposited under accession number ATCC 98337;

^"(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337;

(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity, the fragment comprising the amino acid sequence from amino acid 133 to amino acid 142 of SEQ ID NO:6;

18. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:6;

(b) the amino acid sequence of SEQ ID NO:6 from amino acid 61 to amino acid 256;

(c) fragments of the amino acid sequence of SEQ ID NO:6 comprising the amino acid sequence from amino acid 133 to amino acid 142 of SEQ ID NO:6; and

(d) the amino acid sequence encoded by the cDNA insert of clone CC198_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

19. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:5.

20. An isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 21 to nucleotide 674;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7 from nucleotide 1164 to nucleotide 1465;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CJ317_4 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

21. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:8; (b) fragments of the amino acid sequence of SEQ ID NO:8 comprising the amino acid sequence from amino acid 104 to amino acid 113 of-SEQ ID NO:8; and

^" -(c) the amino acid sequence encoded by the cDNA insert of clone CJ317_4 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

22. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:7.

23. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 951 to nucleotide 1037;

(c) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CS319_1 deposited under accession number ATCC 98337;

(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337;

(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CS319_1 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;

(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 9 to amino acid 18 of SEQ ID NO:10;

(i) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(f) above;

(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ; and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h).

24. ^" - A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 10;

(b) fragments of the amino acid sequence of SEQ ID NO:10 comprising the amino acid sequence from amino acid 9 to amino acid 18 of SEQ ID NO:10; and

(c) the amino acid sequence encoded by the cDNA insert of clone CS319_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

25. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:9 and SEQ ID NO:ll.

26. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:12;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:13; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 13 having biological activity, the fragment comprising the amino acid sequence from amino acid 171 to amino acid 180 of SEQ Il NO:13;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

27. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO:13;

(b) fragments of the amino acid sequence of SEQ ID NO:13 comprising the amino acid sequence from amino acid 171 to amino acid 180 of SEQ ID NO:13; and

(c) the amino acid sequence encoded by the cDNA insert of clone DL504_3 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

28. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:12.

29. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:14;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:14 from nucleotide 1599 to nucleotide 2543;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:14 from nucleotide 174 to nucleotide 396;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DN747J7 deposited under accession number ATCC 98337; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DN747_7 deposited under accession number ATCC 98337;

(f) a polynucleotide comprising Ihe nucleotide sequence of the mature protein -coding sequence of clone DN747_7 deposited under accession number ATCC 98337;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN747_7 deposited under accession number ATCC 98337;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:15 having biological activity, the fragment comprising the amino acid sequence from amino acid 152 to amino acid 161 of SEQ ID NO:15;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

30. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 15;

(b) fragments of the amino acid sequence of SEQ ID NO: 15 comprising the amino acid sequence from amino acid 152 to amino acid 161 of SEQ ID NO:15; and

(c) the amino acid sequence encoded by the cDNA insert of clone DN747J7 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

31. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:14.

32. An isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:16;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 16 from nucleotide 4 to nucleotide 1224;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:17;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:17 having biological activity, the fragment comprising the amino acid sequence from amino acid 198 to amino acid 207 of SEQ ID NO:17;

(j) a polynucleotide which is an allelic variant of a polynucleotide of

(a)-(g) above;

33. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 17;

^" -(d) the amino acid sequence encoded by the cDNA insert of clone DU123_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

34. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:16.

35. An isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18 from nucleotide 233 to nucleotide 370;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18 from nucleotide 293 to nucleotide 370;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:18 from nucleotide 1 to nucleotide 361;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone FB78_1 deposited under accession number ATCC 98337;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337;

(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:19;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:19 having biological activity, the fragment comprising the amino acid sequence from amino acid 18 to amino acid 27 of SEQ ID NO:19; (k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

(1) a polynucleotide which encodes a species homologue of the protein of (i) of {j) above ; and

36. A protein comprising an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of SEQ ID NO: 19;

(b) the amino acid sequence of SEQ ID NO:19 from amino acid 1 to amino acid 43;

(d) the amino acid sequence encoded by the cDNA insert of clone FB78_1 deposited under accession number ATCC 98337; the protein being substantially free from other mammalian proteins.

37. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:18.