EP0972026A2

EP0972026A2 - Secreted proteins and polynucleotides encoding them

Info

Publication number: EP0972026A2
Application number: EP98903573A
Authority: EP
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 LLC
Current assignee: Genetics Institute LLC
Priority date: 1997-01-21
Filing date: 1998-01-21
Publication date: 2000-01-19
Also published as: JP2002512507A; AU6031398A; WO1998031802A3; CA2278153A1; WO1998031802A2

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 the following applications: Ser. No. 60/XXX,XXX (converted to a provisional application from non-provisional application Ser. No.08/786,161), filed January 21, 1997; and Ser. No. 08/877,035, filed June 16, 1997, which is a continuation-in-part of Ser. No. 60/XXX,XXX (converted to a provisional application from non-provisional application Ser. No. 08/786,161), filed January 21, 1997; all of which are 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 101 to nucleotide 428;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:l from nucleotide 325 to nucleotide 461;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone AA365_1 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone AA365_ 1 deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone AA365_1 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone AA365_1 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity;

(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:l from nucleotide 101 to nucleotide 428; the nucleotide sequence of SEQ ID NO:l from nucleotide 325 to nucleotide 461; the nucleotide sequence of the full-length protein coding sequence of clone AA365_1 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone AA365_1 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone AA365_1 deposited under accession number ATCC 98296. 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 76 to amino acid 109. Other embodiments provide the gene corresponding to the cDN A 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 76 to amino acid 109;

(c) fragments of the amino acid sequence of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone AA365_1 deposited under accession number ATCC 98296; 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 76 to amino acid 109.

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 245 to nucleotide 421; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:3 from nucleotide 413 to nucleotide 421;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BL67_2 deposited under accession number ATCC 98296; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BL67_2 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296;

(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;

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

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID

NO:3 from nucleotide 245 to nucleotide 421; the nucleotide sequence of SEQ ID NO:3 from nucleotide 413 to nucleotide 421; the nucleotide sequence of the full-length protein coding sequence of clone BL67_2 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone BL67_2 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296. 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 1 to amino acid 20.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ

ID NO:3 or SEQ ID NO:5. 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:4; (b) the amino acid sequence of SEQ ID NO:4 from amino acid 1 to amino acid 20;

(c) fragments of the amino acid sequence of SEQ ID NO:4; and

(d) the amino acid sequence encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296; 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 1 to amino acid 20.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:6;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:6 from nucleotide 292 to nucleotide 468; (c) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BN130_1 deposited under accession number ATCC 98296;

(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BN130_1 deposited under accession number ATCC 98296; (e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BN130_1 deposited under accession number

ATCC 98296;

(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BN130_1 deposited under accession number ATCC 98296; (g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:7;

(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:7 having biological activity;

(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:6 from nucleotide 292 to nucleotide 468; the nucleotide sequence of the full-length protein coding sequence of clone BN130_1 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone BN130_1 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BN130_1 deposited under accession number ATCC 98296.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:6 or SEQ ID NO:8. 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: 7;

(b) fragments of the amino acid sequence of SEQ ID NO:7; and (c) the amino acid sequence encoded by the cDNA insert of clone

BN130_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:7.

(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 137 to nucleotide 2146; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:9 from nucleotide 218 to nucleotide 2146;

(d) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 189 to nucleotide 397;

(e) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CG99_2 deposited under accession number ATCC 98296;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296; (g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CG99_2 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296;

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

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 10 having biological activity; (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:9 from nucleotide 137 to nucleotide 2146; the nucleotide sequence of SEQ ID NO:9 from nucleotide 218 to nucleotide 2146; the nucleotide sequence of SEQ ID NO:9 from nucleotide 189 to nucleotide 397; the nucleotide sequence of the full-length protein coding sequence of clone CG99_2 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone CG99_2 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10 from amino acid 50 to amino acid 87.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:9. 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:10; (b) the amino acid sequence of SEQ ID NO:10 from amino acid 50 to amino acid 87;

(c) fragments of the amino acid sequence of SEQ ID NO:10; and

(d) the amino acid sequence encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:10 or the amino acid sequence of SEQ ID NO:10 from amino acid 50 to amino acid 87.

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

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 from nucleotide 123 to nucleotide 1247; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO: 11 from nucleotide 1 to nucleotide 419;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CO618_l deposited under accession number ATCC 98296; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CO618_l deposited under accession number

ATCC 98296; (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:12 having biological activity;

(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:ll from nucleotide 123 to nucleotide 1247; the nucleotide sequence of SEQ ID NO:ll from nucleotide 1 to nucleotide 419; the nucleotide sequence of the full-length protein coding sequence of clone CO618_l deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone CO618_l deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 99.

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

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:12; (b) the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 99;

(c) fragments of the amino acid sequence of SEQ ID NO: 12; and

(d) the amino acid sequence encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:12 or the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 99.

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:13;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13 from nucleotide 208 to nucleotide 402; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13 from nucleotide 184 to nucleotide 300;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CO629_2 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CO629_2 deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CO629_2 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CO629_2 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 14 having biological activity;

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

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:13 from nucleotide 208 to nucleotide 402; the nucleotide sequence of SEQ ID NO:13 from nucleotide 184 to nucleotide 300; the nucleotide sequence of the full-length protein coding sequence of clone CO629_2 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone CO629_2 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CO629_2 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14 from amino acid 1 to amino acid 31. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:13.

(a) the amino acid sequence of SEQ ID NO:14;

(b) the amino acid sequence of SEQ ID NO:14 from amino acid 1 to amino acid 31;

(c) fragments of the amino acid sequence of SEQ ID NO: 14; and (d) the amino acid sequence encoded by the cDNA insert of clone

CO629_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:14 or the amino acid sequence of SEQ ID NO: 14 from amino acid 1 to amino acid 31. 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:15;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15 from nucleotide 94 to nucleotide 1059;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15 from nucleotide 1 to nucleotide 387;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CT645_1 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CT645_1 deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CT645_1 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CT645_1 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16 having biological activity;

(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

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:15 from nucleotide 94 to nucleotide 1059; the nucleotide sequence of SEQ ID NO:15 from nucleotide 1 to nucleotide 387; the nucleotide sequence of the full-length protein coding sequence of clone CT645_1 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone CT645_1 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CT645_1 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16 from amino acid 1 to amino acid

98. Other embodiments provide the gene corresponding to the cDNA sequence of SEQ

ID NO:15.

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: 16;

(b) the amino acid sequence of SEQ ID NO: 16 from amino acid 1 to amino acid 98;

(c) fragments of the amino acid sequence of SEQ ID NO: 16; and

(d) the amino acid sequence encoded by the cDNA insert of clone CT645_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:16 or the amino acid sequence of SEQ ID NO: 16 from amino acid 1 to amino acid 98. 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:17; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:17 from nucleotide 640 to nucleotide 1029;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 17 from nucleotide 611 to nucleotide 814;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CW383_1 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CW383_1 deposited under accession number ATCC 98296;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CW383_1 deposited under accession number

ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CW383_1 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:18 having biological activity;

(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:17 from nucleotide 640 to nucleotide 1029; the nucleotide sequence of SEQ ID NO:17 from nucleotide 611 to nucleotide 814; the nucleotide sequence of the full-length protein coding sequence of clone CW383_1 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone CW383_ 1 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CW383_1 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18 from amino acid 1 to amino acid 63.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:17.

(a) the amino acid sequence of SEQ ID NO: 18;

(b) the amino acid sequence of SEQ ID NO:18 from amino acid 1 to amino acid 63;

(c) fragments of the amino acid sequence of SEQ ID NO:18; and (d) the amino acid sequence encoded by the cDNA insert of clone

CW383_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO: 18 or the amino acid sequence of SEQ ID NO:18 from amino acid 1 to amino acid 63. 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:19;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19 from nucleotide 509 to nucleotide 718;

(c) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DN167_1 deposited under accession number ATCC 98296;

(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296;

(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DN167_1 deposited under accession number ATCC 98296; (f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:20 having biological activity;

(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: 19 from nucleotide 509 to nucleotide 718; the nucleotide sequence of the full-length protein coding sequence of clone DN167_1 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone DN167_1 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 65.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO:19. 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:20;

(b) the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 65;

(c) fragments of the amino acid sequence of SEQ ID NO:20; and

(d) the amino acid sequence encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:20 or the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 65.

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:21;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:21 from nucleotide 177 to nucleotide 470; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID

NO:21 from nucleotide 237 to nucleotide 470;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296; (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296; (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296; (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:22;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:22 having biological activity;

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

(a)-(g) above;

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID NO:21 from nucleotide 177 to nucleotide 470; the nucleotide sequence of SEQ ID NO:21 from nucleotide 237 to nucleotide 470; the nucleotide sequence of the full-length protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296; or the nucleotide sequence of the mature protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:22 from amino acid 1 to amino acid 94.

Other embodiments provide the gene corresponding to the cDNA sequence of SEQ ID NO.21.

(a) the amino acid sequence of SEQ ID NO:22; (b) the amino acid sequence of SEQ ID NO:22 from amino acid 1 to amino acid 94;

(c) fragments of the amino acid sequence of SEQ ID NO:22; and

(d) the amino acid sequence encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:22 or the amino acid sequence of SEQ ID NO:22 from amino acid 1 to amino acid 94.

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.

Methods 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 "AA365 1"

A polynucleotide of the present invention has been identified as clone "AA365_1". AA365_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. AA365_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "AA365_1 protein").

The nucleotide sequence of AA365_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 AA365_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:2. Another possible reading frame and predicted amino acid sequence encoded by AA365_1 is encoded by base pairs 157-610 of SEQ ID NO:l and is reported in SEQ ID NO:33. Amino acids 9 to 21 of SEQ ID NO:33 are a predicted leader /signal sequence, with the predicted mature amino acid sequence beginning at amino acid 22 of SEQ ID NO:33, or are a transmembrane domain.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone AA365_1 should be approximately 700 bp.

The nucleotide sequence disclosed herein for AA365_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. AA365_1 demonstrated at least some similarity with sequences identified as AA019239 (ze56c09.sl Soares retina N2b4HR Homo sapiens cDNA clone 362992 3'). Based upon sequence similarity, AA365_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts two potential transmembrane domains within the AA365_1 protein sequence centered around amino acids 33 and 73 of SEQ ID NO:2, respectively; amino acids 14 to 26 are also a possible leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 27 of SEQ ID NO:2.

Clone "BL67 2"

A polynucleotide of the present invention has been identified as clone "BL67_2". BL67_2 was isolated from a human adult testes 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. BL67_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BL67_2 protein").

The nucleotide sequence of the 5' portion of BL67_2 as presently determined is reported in SEQ ID NO:3. What applicants presently believe is the proper reading frame for the coding region is indicated in SEQ ID NO:4. The predicted amino acid sequence of the BL67_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:4. Amino acids 44 to 56 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 57, or are a transmembrane domain. Additional nucleotide sequence from the 3' portion of BL67_2, including the polyA tail, is reported in SEQ ID NO:5.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone BL67_2 should be approximately 970 bp.

The nucleotide sequence disclosed herein for BL67_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. No hits were found in the database.

Clone "BN130 1"

A polynucleotide of the present invention has been identified as clone "BN130_1". BN130_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. BN130_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BN130_1 protein").

The nucleotide sequence of the 5' portion of BN130_1 as presently determined is reported in SEQ ID NO:6. What applicants presently believe is the proper reading frame for the coding region is indicated in SEQ ID NO:7. The predicted amino acid sequence of the BN130_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:7. Additional nucleotide sequence from the 3' portion of BN130_1, including the polyA tail, is reported in SEQ ID NO:8.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone BN130_1 should be approximately 1200 bp. The nucleotide sequence disclosed herein for BN130_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BN130_1 demonstrated at least some similarity with sequences identified as AA023887 (mh93g07.rl Soares mouse placenta 4NbMP13.5 14.5 Mus museums cDNA clone 458556 5'), T99892 (ye68f02.rl Homo sapiens cDNA clone 122907 5'), U65410 (Human Mad2 (hsMAD2) mRNA, complete eds), and Z13859 (H. sapiens partial cDNA sequence; clone 81C04; strand (-), single read). The predicted amino acid sequence disclosed herein for BN130_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BN130_1 protein demonstrated at least some similarity to sequences identified as D37934 (Rat mRNA for 5E5 antigen, complete eds). Based upon sequence similarity, BN130_1 proteins and each similar protein or peptide may share at least some activity.

Clone "CG99 2" A polynucleotide of the present invention has been identified as clone "CG99_2".

CG99_2 was isolated from a human adult testes 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. CG99_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CG99_2 protein").

The nucleotide sequence of CG99_2 as presently determined is reported in SEQ ID NO:9. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CG99_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:10. Amino acids 15 to 27 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 28, or are a transmembrane domain.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone CG99_2 should be approximately 2400 bp. The nucleotide sequence disclosed herein for CG99_2 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CG99_2 demonstrated at least some similarity with sequences identified as L03813 Rattus norvegicus neurotrophin-3 receptor (trkC) mRNA, complete eds). Based upon sequence similarity, CG99_2 proteins and each similar protein or peptide may share at least some activity.

Clone "CO618 1" A polynucleotide of the present invention has been identified as clone "CO618_l".

CO618_l 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. CO618_l is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CO618_l protein").

The nucleotide sequence of CO618_l as presently determined is reported in SEQ ID NO:ll. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CO618_l protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:12.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone CO618_l should be approximately 1750 bp.

The nucleotide sequence disclosed herein for CO618_l was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CO618_l demonstrated at least some similarity with sequences identified as D87074 (Human male bone marrow myeloblast mRNA for KIAA0237 protein, complete eds), H34957 (EST110575 Rattus sp. cDNA), and T33172 (EST56933 Homo sapiens cDNA 3' end similar to None). The predicted amino acid sequence disclosed herein for CO618_l was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CO618_l protein demonstrated at least some similarity to sequences identified as AF007836 (rab3 effector [Rattus norvegicus]), D87074 (KIAA0237 protein; similar to a C.elegans protein encoded in cosmid T10A3(U41035) [Homo sapiens]), and D90195 (polyprotein [Japanese encephalitis virus]). CO618_l protein showed at least some similarity to rat Rim, a putative Rab3 effector. Rim is composed of an amino-terminal zinc-finger motif and carboxy-terminal PDZ and C2 domains. It binds only to GTP (but not to GDP)-complexed Rab3, and interacts with no other Rab protein. Rim is localized to presynaptic active zones in conventional synapses, and to presynaptic ribbons in ribbon synapses (Wang et al, 1997, Nature 388(6642): 593-598, incorporated by reference herein). Based upon sequence similarity, CO618_l proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts a potential transmembrane domain within the CO618_l protein sequence centered around amino acid 350 of SEQ ID NO:12.

Clone "CO629 2"

A polynucleotide of the present invention has been identified as clone "CO629_2". CO629_2 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. CO629_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CO629_2 protein").

The nucleotide sequence of CO629_2 as presently determined is reported in SEQ ID NO:13. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CO629_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:14. Another possible reading frame and predicted amino acid sequence encoded by CO629_2 is encoded by basepairs 417 to 632 of SEQ ID NO:13 and is reported in SEQ ID NO:34; this alternative open reading frame could be joined to the reading frame reported in SEQ ID NO: 14 if an insertion or deletion resulting in a frameshift was made in the sequence of SEQ ID NO:13. The TopPredll computer program predicts a potential transmembrane domains within the alternative CO629_2 protein sequence centered around amino acid 50 of SEQ ID NO:34.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone CO629_2 should be approximately 1800 bp.

The nucleotide sequence disclosed herein for CO629_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CO629_2 demonstrated at least some similarity with sequences identified as U78857 (Rattus norvegicus calcium calmodulin dependent kinase CPG16 (cpglό) mRNA, complete eds) and W77588 (me67bll.rl Soares mouse embryo NbME13.5

14.5 Mus musculus cDNA). The predicted amino acid sequence disclosed herein for CO629_2 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CO629_2 protein demonstrated at least some similarity to sequences identified as U78857 (calcium calmodulin dependent kinase CPG16 [Rattus norvegicus]). Based upon sequence similarity, C0629_2 proteins and each similar protein or peptide may share at least some activity.

Clone "CT645 1" A polynucleotide of the present invention has been identified as clone "CT645_1".

CT645_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. CT645_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CT645_1 protein").

The nucleotide sequence of CT645_1 as presently determined is reported in SEQ ID NO:15. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CT645_ 1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:16.

The EcoRI/Notl restriction fragment obtainable from the deposit containing clone CT645_1 should be approximately 2560 bp.

The nucleotide sequence disclosed herein for CT645_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CT645_1 demonstrated at least some similarity with sequences identified as AA430737 (zw20gl2.rl Soares ovary tumor NbHOT Homo sapiens cDNA clone 769894 5'), H34543 (EST111568 Rattus sp. cDNA 5' end), and N34881 (yy46gl2.sl Homo sapiens cDNA clone 276646 3' similar to contains element MER37 repetitive element). Based upon sequence similarity, CT645_1 proteins and each similar protein or peptide may share at least some activity. The TopPredll computer program predicts two potential transmembrane domains within the CT645_1 protein sequence centered around amino acids 75 and 275 of SEQ ID NO:16, respectively; amino acids 92 to 104 of SEQ ID NO:16 are a possible leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 105. The nucleotide sequence of CT645_1 indicates that it may contain an Alu repetitive element.

Clone "CW383 1"

A polynucleotide of the present invention has been identified as clone "CW383_1". CW383_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. CW383_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CW383_1 protein").

The nucleotide sequence of CW383_1 as presently determined is reported in SEQ ID NO:17. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the CW383_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:18. The EcoRI/Notl restriction fragment obtainable from the deposit containing clone

CW383_1 should be approximately 1000 bp.

The nucleotide sequence disclosed herein for CW383_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CW383_1 demonstrated at least some similarity with sequences identified as AA037470 (zk36g04.sl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 484950 3'), AA037478 (zk36h05.sl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 484953 3'), AA088729 (zl89c09.rl Stratagene colon (#937204) Homo sapiens cDNA clone 511792 5'), AA098551 (mkl8fl2.rl Soares mouse p3NMF19.5 Mus musculus cDNA clone 493295 5'), R75968 (yi62d07.rl Homo sapiens cDNA clone 143821 5'), and X85681 (H.sapiens mRNA for expressed sequence tag, clone CAM tESTlFό (B)). Based upon sequence similarity, CW383_1 proteins and each similar protein or peptide may share at least some activity.

Clone "DN167 1" A polynucleotide of the present invention has been identified as clone "DN167_1".

DN167_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. DN167_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DN167_1 protein").

The nucleotide sequence of DN167_1 as presently determined is reported in SEQ ID NO:19. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DN167_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:20.

The EcoRI /Notl restriction fragment obtainable from the deposit containing clone DN167_1 should be approximately 850 bp. The nucleotide sequence disclosed herein for DN167_1 was searched against the

GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DN167_1 demonstrated at least some similarity with sequences identified as AA405121 (zu65d01.rl Soares testis NHT Homo sapiens cDNA clone 742849 5'), AC000045 (00064; HTGS phase 3, complete sequence), R51018 (yg71b02.sl Homo sapiens cDNA clone 38639 3' similar to contains Alu repetitive elemen contains MSRl repetitive element), and Z70233 (Human DNA sequence from cosmid V870H8, between markers DXS366 and DXS87 on chromosome X contains ESTs). Based upon sequence similarity, DN167_1 proteins and each similar protein or peptide may share at least some activity. The nucleotide sequence of DN167_1 indicates that it may contain an Alu repetitive element.

Clone "DN711 2"

A polynucleotide of the present invention has been identified as clone "DN711_2". DN711_2 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. DN711_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DN711_2 protein"). The nucleotide sequence of DN711_2 as presently determined is reported in SEQ

ID NO:21. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DN711_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:22. 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/Notl restriction fragment obtainable from the deposit containing clone DN711_2 should be approximately 700 bp.

The nucleotide sequence disclosed herein for DN711_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DN711_2 demonstrated at least some similarity with sequences identified as AA127018 (zl22a03.sl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 502636 3'), AA128221 (zl22a03.rl Soares pregnant uterus NbHPU Homo sapiens cDNA clone 502636 5'), AA444139 (zvδlfOl.rl Soares testis NHT Homo sapiens cDNA clone 757177 5'), AA460975 (zx63cll.sl Soares total fetus Nb2HF8 9w Homo sapiens), D62671 (Human aorta cDNA 5'-end GEN-312F07), D79837 (Human aorta cDNA 5'-end GEN-335E12), D79860 (Human aorta cDNA 5'-end GEN-342C02), R24595 (yg36h06.rl Homo sapiens cDNA clone 34631 5'), and R40285 (yf80h03.sl Homo sapiens cDNA clone). Based upon sequence similarity, DN711_2 proteins and each similar protein or peptide may share at least some activity.

Deposit of Clones

Clones AA365_1, BL67_2, BN130_1, CG99_2, CO618_l, CO629_2, CT645_1, CW383_1, DN167_1, and DN711_2 were deposited on January 17, 1997 with the American Type Culture Collection as an original deposit under the Budapest Treaty and were given the accession number ATCC 98296, 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/Notl digestion (5' site, EcoRI; 3' site, Notl) 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 pEDόdpcl 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 Notl. However, Notl 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 AA365_1 SEQ ID NO.23

BL67_2 SEQ ID NO:24

BN130_1 SEQ ID NO:25

CG99_2 SEQ ID NO:26

CO618_l SEQ ID NO:27 CO629_2 SEQ ID NO:28

CT645_1 SEQ ID NO:29

CW383_1 SEQ ID NO:30

DN167_1 SEQ ID NO:31

DN711_2 SEQ ID NO:32

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-O-(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 transposable 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. Proteins 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.

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 region(s) of the hybridizmg polynucleotides When hybridizing 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 (lxSSC 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 hybndization 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 + 166(log,₀[Na⁺]) + 0.41 (%G+C) - (600/N), where N is the number of bases m the hybrid, and [Na⁺] is the concentration of sodium ions in the hybridization buffer ([Na⁺] for lxSSC = 0.165 M). Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, 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 typhimurium, 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/lpr/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 of 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 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. The activity of a protein of the invention may, among other means, be measured by the following methods:

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 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); 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 have 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. WO95/ 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 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.

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

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 detected 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, TNFl, 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-oc 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, Vi ki Agostino, Michael J.

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

(iii) NUMBER OF SEQUENCES: 34

(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: Patentln 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: 633 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS : double

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

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

CTCTTACCCC TCCTCCTGTT TCATGGACAT TTCAGTGTTG CCAGTGAAAA AGTAATGGTT 60

TTAATTTGGT CCTTATTTTT AACCTGCCCC TGAGACTTAT ATGCTTGTTT ATACCATGTA 120

CGTAGTGTGT GATTGTATGT GTTTGTATTT GTCCACATGT CCCAAAACAT GGGCTGTTAC 180

TTCCTTTTTC TATCTTGGTT TCCTTATTCC CACCCTTCTT TTTCCACCCA GGTATCTGGA 240

CAGGAAGACT TCTCCCATCA GCTTTACCAG AGGAAGCTGC AGGCCCCACT GTGGCCCAGC 300

TCCCTGGGCA TCACTGATTG CTGTCAGTAT GTCACCTCCT GTCACCCCAA GAGATCAGAG 360

AGACGCAAGT ATGGCCGAGA CTTCCTGCTA CGTTTCCGCT TCTGCAGCAT CGCTTGTCAG 420

CGACCAGTAG GACTGGTCCT TATGGAAGGA GTGACAGATA CTAAGCCAGA GCGACCTGCG 480

GGTTGGGCTG AGTCTGTCCT TGAGGAAGAT GCATCGGAGC TTGAGCCTGC CTTCTCCAGG 540

ACTGTAGGTA CCATCCAGCA CTGCCTCCAC CTGACGTCAG TATATACCCC CTCTACCCCC 600

TCTAGAGCCA AAAAAAAAAA AAAAAAAAAA AAA 633 (2) INFORMATION FOR SEQ ID NO : 2 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 109 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Met Leu Val Tyr Thr Met Tyr Val Val Cys Asp Cys Met Cys Leu Tyr 1 5 10 15

Leu Ser Thr Cys Pro Lys Thr Trp Ala Val Thr Ser Phe Phe Tyr Leu 20 25 30

Gly Phe Leu lie Pro Thr Leu Leu Phe Pro Pro Arg Tyr Leu Asp Arg 35 40 45

Lys Thr Ser Pro lie Ser Phe Thr Arg Gly Ser Cys Arg Pro His Cys 50 55 60 Gly Pro Ala Pro Trp Ala Ser Leu He Ala Val Ser Met Ser Pro Pro 65 70 75 80

Val Thr Pro Arg Asp Gin Arg Asp Ala Ser Met Ala Glu Thr Ser Cys 85 90 95

Tyr Val Ser Ala Ser Ala Ala Ser Leu Val Ser Asp Gin 100 105

(2) INFORMATION FOR SEQ ID NO : 3 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 421 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

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

GCGGCCGCAG GTCTAGAATT CAATCGGGGA AGTGGGCTGT CCAGCTGATG TGCAGCCTCA 60

AAAGAGACCC CAAGCCAGAC CCACCCACGA AAGGTACCCC TGAATTCTGG ACTCATAAAA 120

TCTCTTAGAT AATACATATT TGTTGTTTTA ACCCACTCAG TTTTGCATTG GTTACACAGC 180

AATAGAAAAT AAAGGAGGCA GATGAAATTG ATGGAGAACA ATTGCAACGA AAACAGAATA 240

CACAATGCAC GAGCCTGTGT CAGGAATGAC AGTGCATTCC ACGGAAGAGT TGCACAGAGA 300

GAGACCAGTT ACAAGATGCC ATCAGGATAA CGGGAGCTGC GCTGGGCAGC TGGGCCTTCC 360

GACATCCTCC CTGGTTGCGG TGGCAGTGGT GGTGTCTGCT CCAAGTCAAG GCTGGCAGAG 420

A 421 (2) INFORMATION FOR SEQ ID NO : 4 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 59 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 4 : Met His Glu Pro Val Ser Gly Met Thr Val His Ser Thr Glu Glu Leu 1 5 10 15

His Arg Glu Arg Pro Val Thr Arg Cys His Gin Asp Asn Gly Ser Cys 20 25 30

Ala Gly Gin Leu Gly Leu Pro Thr Ser Ser Leu Val Ala Val Ala Val 35 40 45

Val Val Ser Ala Pro Ser Gin Gly Trp Gin Arg 50 55

(2) INFORMATION FOR SEQ ID NO : 5 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 393 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

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

TCTGGAAACA CAAACCTCAC TGAGTAACAA AGAAACAGTA TTTGAGGAAA GCCAATGTGA 60

TGAAAGAGAG ACGCCAGACT GGAAGGAGGG ACGGTCATCT GAGAACATAC AAAGCAAGTG 120

GCAAGCGTTA GTTTACACGT CCAAGGTGAC CGTCACATGC TCTGTCCAGG CAAGAGAGGC 180

AAGTGGACAC CCTGAGAAGG AAAAATGTTG TCAAAATAAA AACCTAATAT GTGAACCTAG 240

GAGTAAAATG GACACAGCTT GCGAGCAAAG AGACAGTGAG AAGATGAAGC CAAGGCACTC 300

TCCAGAATGC AGGTGACGGG AAAAAGAGAC AGTAACAGAT CCAAAGAGGT CCAGAAGGAA 360

TGACCATGGA GCGGAAAGAA AGAAAAAAAA AAA 393 (2) INFORMATION FOR SEQ ID NO : 6 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 468 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 6 : GCTCCGGTGG CTGCGCAGCG TCGGCGGGGA GCGCGGGCTG CGGAGAGGCG GGCCGGGCCA 60

AGCGGAGCCG AGCGAGCGGG AGCGCGGCGT CCGGGAGGCG GCGGAGACGC GGGGCTCGGA 120

GGGTCAGCCT CTTATCGTAG CAGGTCTCCT CGGCACGCCC CCCTTGTTTC GCCCCACGGC 180

CAAGCCCGCC GCGGGCCGGC GTGCGCTGGT CACTGAGGCC CAGGTCGCCG CCGCGGCGCG 240

TTTTTGAAAT CATGAATCCT GTTTATAGTC CTGGATCTTC TGGGGTTCCC TATGCAAATG 300

CCAAAGGAAT TGGTTATCCA GCTGGTTTTC CCATGGGCTA TGCAGCAGCA GCTCCCTGCC 360

TATTCTCCTA ACATGTATCC TGGAGCGAAT CCTACCTTCC AAACAGGTTA CACTCCTGGC 420

ACACCTTACA AAGTGTCCTG TTCCCCCACC AGCGGGGCTG TGCCACCG 468 (2) INFORMATION FOR SEQ ID NO : 7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 57 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO : 7 :

Met Pro Lys Glu Leu Val He Gin Leu Val Phe Pro Trp Ala Met Gin 1 5 10 15

Gin Gin Leu Pro Ala Tyr Ser Pro Asn Met Tyr Pro Gly Ala Asn Pro 20 25 30

Thr Phe Gin Thr Gly Tyr Thr Pro Gly Thr Pro Tyr Lys Val Ser Cys 35 40 45

Ser Pro Thr Ser Gly Ala Val Pro Pro 50 55

(2) INFORMATION FOR SEQ ID NO : 8 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 366 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 8 :

AAGTACCGGG CCCCAGGAAC GCCCACTTAC AGCTATGTGC CCCCTCAGTG GTGATCACCT 60

GCAAATGTTT GAGGACGGAG CTGTGCAGTC ACATTATTGG GGATTCCACA GCTGGTGCTG 120

CAGGCCTTGC GCCTCCAACC AGGACTTTCT TCTTAATGCT CTCGACACTT AGCTAAACAC 180

GACTATATCC CGGCCCAGCA GGCCCCAGCG CCGTTAGTCT CCAGCTGACT CTGTGGGTTG 240

GTCTTAAAGC AAATTCTGTT TTGTGGACTG CCTGGCAATT TTTTAGCTAA CTGTAATGAT 300

AAAAAGGGAG TATTAATCTA TTCTGAATCA TATCTAGTTG AATGCATGTT TAAAAAAAAA 360

AAAAAA 366 (2) INFORMATION FOR SEQ ID NO : 9 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2423 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

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

AGGGCCCGCC GCTGGGATGC CGAGCGCCCG CGCCGCCGCT GCCTCTGTCC TCCGCGCGCT 60

GCTCAGCTGA AGGCGCACAG GATTCAATTA CTGGACTTGT CAACTCTGCC AGTGTACGTG 120

CCATTTCTCT TCCACTATGA GAGGACCGAT TGTATTGCAC ATTTGTCTGG CTTTCTGTAG 180

CCTTCTGCTT TTCAGCGTTG CCACACAATG TCTGGCCTTC CCCAAAATAG AAAGGAGGAG 240

GGAGATAGCA CATGTTCATG CGGAAAAAGG GCAGTCCGAT AAGATGAACA CCGATGACCT 300

AGAAAATAGC TCTGTTACCT CAAAGCAGAC TCCCCAACTG GTGGTCTCTG AAGATCCAAT 360

GATGATGTCA GCAGTACCAT CGGCAACATC ATTAAATAAA GCATTCTCGA TTAACAAAGA 420

AATCCAGCCT GGACAAGCTG GGCTCATGCA AACAGAACGC CCTGGTGTTT CCACACCTAC 480

TGAGTCAGGT GTCCCCTCAG CTGAAGAAGT ATTTGGTTCC AGCCAGCCAG AGAGAATATC 540

TCCTGAAAGT GGACTTGCCA AGGCCATGTT AACCATTGCT ATCACTGCGA CTCCTTCTCT 600

GACTGTTGAT GAAAAGGAGG AACTCCTTAC AAGCACTAAC TTTCAGCCCA TTGTAGAAGA 660

GATCACAGAA ACCACAAAAG GTTTTCTGAA GTATATGGAT AATCAATCAT TTGCAACTGA 720 AAGTCAGGAA GGAGTTGGTT TGGGACATTC ACCTTCATCC TATGTGAATA CTAAGGAAAT 780

GCTAACCACC AATCCAAAGA CTGAGAAATT TGAAGCAGAC ACAGACCACA GGACAACTTC 840

TTTTCCTGGT GCTGAGTCCA CAGCAGGCAG TGAGCCTGGA AGCCTCACCC CTGATAAGGA 900

GAAGCCTTCG CAGATGACAG CTGATAACAC CCAGGCTGCT GCCACCAAGC AACCACTCGA 960

AACTTCCGAG TACACCCTGA GTGTTGAGCC AGAAACTGAT AGTCTGCTGG GAGCCCCAGA 1020

AGTCACAGTG AGTGTCAGCA CAGCTGTTCC AGCTGCCTCT GCCTTAAGTG ATGAGTGGGA 1080

TGACACCAAA TTAGAGAGTG TAAGCCGGAT AAGGACCCCC AAGCTTGGAG ACAATGAAGA 1140

GACTCAGGTG AGAACGGAGA TGTCTCAGAC AGCACAAGTA AGCCATGAGG GTATGGAAGG 1200

AGGCCAGCCT TGGACAGAGG CTGCACAGGT GGCTCTGGGG CTGCCTGAAG GGGAAACACA 1260

CACGGGCACA GCCCTGCTAA TAGCGCATGG GAATGAGAGA TCACCTGCTT TCACTGATCA 1320

AAGTTCCTTT ACCCCCACAA GTCTGATGGA AGACATGAAA GTTTCCATTG TGAACTTGCT 1380

CCAAAGTACG GGAGACTTCA CGGAATCCAC CAAGGAAAAC GATGCCCTGT TTTTCTTAGA 1440

AACCACTGTT TCTGTCTCTG TATATGAGTC TGAGGCAGAC CAACTGTTGG GAAATACAAT 1500

GAAAGACATC ATCACTCAAG AGATGACAAC AGCTGTTCAA GAGCCAGATG CCACTTTATC 1560

CATGGTGACA CAAGAGCAGG TTGCTACCCT CGAGCTTATC AGAGACAGTG GCAAGACTGA 1620

GGAAGAAAAG GAGGACCCCT CTCCTGTGTC TGACGTTCCT GGTGTTACTC AGCTGTCAAG 1680

AAGATGGGAG CCTCTGGCCA CTACAATTTC AACTACAGTC GTCCCTTTGT CTTTTGAAGT 1740

TACTCCCACT GTGGAAGAAC AAATGGACAC AGTCACAGGG CCAAATGAGG AGTTCACACC 1800

AGTTCTGGGA TCTCCAGTGA CACCTCCTGG AATAATGGTG GGGGAACCCA GCATTTCCCC 1860

TGCACTTCCT GCTTTGGAGG CATCCTCTGA GAGAAGAACT GTTGTTCCAT CTATTACTCG 1920

TGTTAATACA GCTGCCTCAT ATGGCCTGGA CCAACTTGAA TCTGAAGAGA CGGGGTTTCA 1980

CCATGTTGCC CAGGCTCGTC TCAAACTCCT GGGCTTAAGA AGTCTGCCTG CCTCGGCCTC 2040

CCAAAGTGTT GGAATTACAA GTGTGAACTC CTGCACCCAG CCAAGAAAAT ATTTAAACAG 2100

CTGCCTGAAA TGGAAATTGA ATCCAAAACA TTTTGCCACT GGTGTTTGAA GTGGACTATC 2160

TTGTCTC AG CCTTTTTATT TTCTCATGAA TGTACCTTCT AAATATTGTT AAGGCTACAA 2220

TGGAGGAAAT TTTAAATTAT TATATTTACT ATTTTACAGT GAGTGGAAAA TTTGAAATTT 2280

AGTAAAAAGT TTTCAAGAAA TTTTTATAGA TTAATAATGG AGTTTAAAAA GTCATGGCGA 2340

TGAGGCATTT AACAAGGAGA GAACTGAATA AATTAACAGA CATGTTTAAG GTAAAAAAAA 2400 AAAAAAAAAA AAAAAAAAAA AAA 2423

(2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 670 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Met Arg Gly Pro He Val Leu His He Cys Leu Ala Phe Cys Ser Leu 1 5 10 15

Leu Leu Phe Ser Val Ala Thr Gin Cys Leu Ala Phe Pro Lys He Glu 20 25 30

Arg Arg Arg Glu He Ala His Val His Ala Glu Lys Gly Gin Ser Asp 35 40 45

Lys Met Asn Thr Asp Asp Leu Glu Asn Ser Ser Val Thr Ser Lys Gin 50 55 60

Thr Pro Gin Leu Val Val Ser Glu Asp Pro Met Met Met Ser Ala Val 65 70 75 80

Pro Ser Ala Thr Ser Leu Asn Lys Ala Phe Ser He Asn Lys Glu He 85 90 95

Gin Pro Gly Gin Ala Gly Leu Met Gin Thr Glu Arg Pro Gly Val Ser 100 105 110

Thr Pro Thr Glu Ser Gly Val Pro Ser Ala Glu Glu Val Phe Gly Ser 115 120 125

Ser Gin Pro Glu Arg He Ser Pro Glu Ser Gly Leu Ala Lys Ala Met 130 135 140

Leu Thr He Ala He Thr Ala Thr Pro Ser Leu Thr Val Asp Glu Lys 145 150 155 160

Glu Glu Leu Leu Thr Ser Thr Asn Phe Gin Pro He Val Glu Glu He 165 170 175

Thr Glu Thr Thr Lys Gly Phe Leu Lys Tyr Met Asp Asn Gin Ser Phe 180 185 190

Ala Thr Glu Ser Gin Glu Gly Val Gly Leu Gly His Ser Pro Ser Ser 195 200 205 Tyr Val Asn Thr Lys Glu Met Leu Thr Thr Asn Pro Lys Thr Glu Lys 210 215 220

Phe Glu Ala Asp Thr Asp His Arg Thr Thr Ser Phe Pro Gly Ala Glu 225 230 235 240

Ser Thr Ala Gly Ser Glu Pro Gly Ser Leu Thr Pro Asp Lys Glu Lys 245 250 255

Pro Ser Gin Met Thr Ala Asp Asn Thr Gin Ala Ala Ala Thr Lys Gin 260 265 270

Pro Leu Glu Thr Ser Glu Tyr Thr Leu Ser Val Glu Pro Glu Thr Asp 275 280 285

Ser Leu Leu Gly Ala Pro Glu Val Thr Val Ser Val Ser Thr Ala Val 290 295 300

Pro Ala Ala Ser Ala Leu Ser Asp Glu Trp Asp Asp Thr Lys Leu Glu 305 310 315 320

Ser Val Ser Arg He Arg Thr Pro Lys Leu Gly Asp Asn Glu Glu Thr 325 330 335

Gin Val Arg Thr Glu Met Ser Gin Thr Ala Gin Val Ser His Glu Gly 340 345 350

Met Glu Gly Gly Gin Pro Trp Thr Glu Ala Ala Gin Val Ala Leu Gly 355 360 365

Leu Pro Glu Gly Glu Thr His Thr Gly Thr Ala Leu Leu He Ala His 370 375 380

Gly Asn Glu Arg Ser Pro Ala Phe Thr Asp Gin Ser Ser Phe Thr Pro 385 390 395 400

Thr Ser Leu Met Glu Asp Met Lys Val Ser He Val Asn Leu Leu Gin 405 410 415

Ser Thr Gly Asp Phe Thr Glu Ser Thr Lys Glu Asn Asp Ala Leu Phe 420 425 430

Phe Leu Glu Thr Thr Val Ser Val Ser Val Tyr Glu Ser Glu Ala Asp 435 440 445

Gin Leu Leu Gly Asn Thr Met Lys Asp He He Thr Gin Glu Met Thr 450 455 460

Thr Ala Val Gin Glu Pro Asp Ala Thr Leu Ser Met Val Thr Gin Glu 465 470 475 480

Gin Val Ala Thr Leu Glu Leu He Arg Asp Ser Gly Lys Thr Glu Glu 485 490 495

Glu Lys Glu Asp Pro Ser Pro Val Ser Asp Val Pro Gly Val Thr Gin 500 505 510

Leu Ser Arg Arg Trp Glu Pro Leu Ala Thr Thr He Ser Thr Thr Val 515 520 525

Val Pro Leu Ser Phe Glu Val Thr Pro Thr Val Glu Glu Gin Met Asp 530 535 540

Thr Val Thr Gly Pro Asn Glu Glu Phe Thr Pro Val Leu Gly Ser Pro 545 550 555 560

Val Thr Pro Pro Gly He Met Val Gly Glu Pro Ser He Ser Pro Ala 565 570 575

Leu Pro Ala Leu Glu Ala Ser Ser Glu Arg Arg Thr Val Val Pro Ser 580 585 590

He Thr Arg Val Asn Thr Ala Ala Ser Tyr Gly Leu Asp Gin Leu Glu 595 600 605

Ser Glu Glu Thr Gly Phe His His Val Ala Gin Ala Arg Leu Lys Leu 610 615 620

Leu Gly Leu Arg Ser Leu Pro Ala Ser Ala Ser Gin Ser Val Gly He 625 630 635 640

Thr Ser Val Asn Ser Cys Thr Gin Pro Arg Lys Tyr Leu Asn Ser Cys 645 650 655

Leu Lys Trp Lys Leu Asn Pro Lys His Phe Ala Thr Gly Val 660 665 670

(2) INFORMATION FOR SEQ ID NO: 11:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1302 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

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

CAGGATGGGA TCCTCATAGA GGGGCAGATA ATGTTTCTAC TAAATCTTCG GACAGTGATG 60

TAAGTGATAT ATCTGCGGTT TCAAGGACTA GTAGTGCTTC TCGTTTCAGC AGCACAAGCT 120

ACATGTCTGT CCAATCAGAA CGCCCAGGAG GAAACAAGAA AATCAGTGTC TTTACATCCA 180

AAATGCAAAG CAGACAAATG GGCATATCAG GGAAGAACAT GACAAAAAGC ACCAGCATCA 240 GTGGAGACAT GTGCTCACTG GAGAAGAATG ATGGCAGCCA GTCTGACACT GCAGTGGGCA 300 CCTTGGGCAC CAGTGGCAAA AAGCGGCGCT CTAGCCTTGG TGCCAAAATG GTAGCTATCG 360 TTGGTCTGTC ACGGAAAAGT CGCAGTGCTT CTCAGCTCAG CCAAACGGAA GCAGGAGGTW 420 AAAAACTAAG GAGCACTGTC CAAAGAAGTA CAGAAACAGG CCTGGCCGTG GAAATGAGGA 480 ACTGGATGAC TCGACAGGCA AGCCGAGAGT CTACAGATGG TAGCATGAAC AGCTACAGCT 540 CAGAAGGAAA TCTGATTTTC CCTGGTGTTC GCTTGGCCTC TGATAGCCAG TTCAGTGATT 600 TCCTGGATGG CCTTGGCCCT GCTCAGCTAG TGGGACGCCA GACTCTGGCA ACACCTGCAA 660 TGGGTGACAT TCAGGTAGGA ATGATGGACA AAAAGGGACA GCTGGAGGTA GAAATCATCC 720 GGGCCCGTGG CCTTGTTGTA AAACCAGGTT CCAAGACACT GCCAGCACCG TATGTAAAAG 780 TGTATCTATT AGATAACGGA GTCTGCATAG CCAAAAAGGG ACAGCTGGAG GTAGAAATCA 840 TCCGGGCCCG TGGCCTTGTT GTAAAACCAG GTTCCAAGAC ACTGCCAGCA CCGTATGTAA 900 AAGTGTATCT ATTAGATAAC GGAGTCTGSA TAGCCAAAAA GAAAACAAAA GTGGCAAGAA 960

AAACGCTGGA ACCCCTTTAC CAGCAGYTAT TATCTTTCGA AGAGAGTCCA CAAGGAAAAG 1020

TTTTACAGAT CATCGTCTGG GGAGATTATG GCCGCATGGA TCACAAATCT TTTATGGGAG 1080

TGGCCCAGAT ACTTTTAGAT GAACTAGAGC TATCCAATAT GGTGATCGGA TGGTTCAAAC 1140

TTTTCCCACC TTCCTCCCTA GTAGATCCAA CCTTGGCCCC TCTGACAAGA AGAGCTTCCC 1200

AATCATCTCT GGAAAGTTCA ACTGGACCTT CTTACTYTCG TTCATAGCAG CTGTAAAAAA 1260

ATTGTTGTCA CAGCAACCAG CGTTACAAAA AAAAAAAAAA AA 1302 (2) INFORMATION FOR SEQ ID NO: 12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 374 amino acids

(B) TYPE: amino acid

( C ) STRANDEDNESS :

( D) TOPOLOGY : linear

(ii) MOLECULE TYPE: protein

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

Met Ser Val Gin Ser Glu Arg Pro Gly Gly Asn Lys Lys He Ser Val 1 5 10 15

Phe Thr Ser Lys Met Gin Ser Arg Gin Met Gly He Ser Gly Lys Asn 20 25 30

Met Thr Lys Ser Thr Ser He Ser Gly Asp Met Cys Ser Leu Glu Lys 35 40 45

Asn Asp Gly Ser Gin Ser Asp Thr Ala Val Gly Thr Leu Gly Thr Ser 50 55 60

Gly Lys Lys Arg Arg Ser Ser Leu Gly Ala Lys Met Val Ala He Val 65 70 75 80

Gly Leu Ser Arg Lys Ser Arg Ser Ala Ser Gin Leu Ser Gin Thr Glu 85 90 95

Ala Gly Gly Xaa Lys Leu Arg Ser Thr Val Gin Arg Ser Thr Glu Thr 100 105 110

Gly Leu Ala Val Glu Met Arg Asn Trp Met Thr Arg Gin Ala Ser Arg 115 120 125

Glu Ser Thr Asp Gly Ser Met Asn Ser Tyr Ser Ser Glu Gly Asn Leu 130 135 140

He Phe Pro Gly Val Arg Leu Ala Ser Asp Ser Gin Phe Ser Asp Phe 145 150 155 160

Leu Asp Gly Leu Gly Pro Ala Gin Leu Val Gly Arg Gin Thr Leu Ala 165 170 175

Thr Pro Ala Met Gly Asp He Gin Val Gly Met Met Asp Lys Lys Gly 180 185 190

Gin Leu Glu Val Glu He He Arg Ala Arg Gly Leu Val Val Lys Pro 195 200 205

Gly Ser Lys Thr Leu Pro Ala Pro Tyr Val Lys Val Tyr Leu Leu Asp 210 215 220

Asn Gly Val Cys He Ala Lys Lys Gly Gin Leu Glu Val Glu He He 225 230 235 240

Arg Ala Arg Gly Leu Val Val Lys Pro Gly Ser Lys Thr Leu Pro Ala 245 250 255

Pro Tyr Val Lys Val Tyr Leu Leu Asp Asn Gly Val Xaa He Ala Lys 260 265 270

Lys Lys Thr Lys Val Ala Arg Lys Thr Leu Glu Pro Leu Tyr Gin Gin 275 280 285

Leu Leu Ser Phe Glu Glu Ser Pro Gin Gly Lys Val Leu Gin He He 290 295 300

Val Trp Gly Asp Tyr Gly Arg Met Asp His Lys Ser Phe Met Gly Val 305 310 315 320 Ala Gin He Leu Leu Asp Glu Leu Glu Leu Ser Asn Met Val He Gly 325 330 335

Trp Phe Lys Leu Phe Pro Pro Ser Ser Leu Val Asp Pro Thr Leu Ala 340 345 350

Pro Leu Thr Arg Arg Ala Ser Gin Ser Ser Leu Glu Ser Ser Thr Gly 355 360 365

Pro Ser Tyr Xaa Arg Ser 370

(2) INFORMATION FOR SEQ ID NO: 13:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1627 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:

GGCCTACTGA ATAACAATAG TTATTTTTGA AAGATCATTG TTTCCCTAAT TCTGTGGCTA 60

AGAGGGGCCC ATTTAGGAAG AGGTCAGGAG GGTGGTTGAG GCGGGGAGGG GGACTCTTAA 120

GGAAAAGACA ACTCAAACGG GAGGATGAAA AAGCATTGTA TTTGATTGTT TTCTTTCCTG 180

TAGGATGATG CCTCCCAGGA GAATAACATG CAAGCTGAGG TGACAGGTAA ACTAAAACAG 240

CACTTTAATA ATGCGCTCCC CAAACAGAAC AGCACTACCA CCGGGGTCTC CGTCATCATG 300

GTGAGTGGAA GGCGGCAGGT CTGGCCTGAC TGCGGAGCCG GCCTTGAAGT TTTTGAATTA 360

GGTAGCCGGG AGCTGCCCTC ACATGGAAGT TGGTGCCTTC CGTAGTCCTA TTTCATATGA 420

AGATTGGCTT GGCATGTGGA GGGCACTCAT TCGGCAACTC CCAGGCTTTG GGCACTGTGT 480

GGAGGGGCTT GTGTAGGGAC CAGCAGGCCT GGTGTGAGGG GTCCAGGCGT CAAGGAGCTC 540

CTGGCTGGGC CCTCTGGGCA GCTGCTTCCA CTCTTGTCTC TGCCTTCTCA TCTAGAGAGA 600

CTCCCAAGCC CTGGAGGGGT GTGTTGTGTT AGGAATTAAC TCCCTGCCTA CCCCAAGGCC 660

TCAGAAATAG ATTATTAGAG ATGTGAATTA TTCTTTGAGA CTTGGGATAA GAAACAGCCA 720

AAGCTAAACA TATTTCAGTT TTAAAAAATC AGTGTTTTAT AAAACACAGT TTGGGGCTTT 780

TAAAGGTACA TAATCAAGGA AAAAAATATA TATTCATTTT TCAGGGTTGG TAACATTTTA 840 TGAGATGTCA GTGACAACGA TGGCCTTATT TTTTTCAGCC TTTTCTTCTT CCAAAATGTT 900 TCTTAAGGCA ACTCTCCTAA ATACATAAAC ACAACAAATT AAAATGAAAA GTGACATGAG 960

AGTAAATGAA TCAAAAGGAA AAAACATTGA ACCAGAGGTG AGGGCAGCAC ACCCGCAGCA 1020

GCTGTCCAGG CCTGAGCCAA TGCAACCCTG GGCGGGAAGG CCAGCTCACC GTGAGCAGGT 1080

AGAAGCCAGC CAGCCACCCA GGCAGGGACC TTGGTTCTCC CCACACACTC CCAGGAGCAG 1140

GGAACAGGGG TGGAGTGGCC TTTCCCAGAG CTGGAGTTGG CTGCAGCAGC TTTCGAATCA 1200

GACCTGCCAA GGTGATGGGC GTCTGAGTTT CACATCTGGG CCCCCCGTGA CCCCACTGAG 1260

TCCTGACAGC TAAGGATGGG CCACCTCCAC AGCTCCGTCA CTCGTACTTG GGACAGGCCT 1320

CTCATCCTCT GGGAAGGTCC TCCTTGTTTC CTACCCAACT AGAAGGGAAA CAGTGGCATA 1380

TTCTCATGGT ACATGGTTGT CTGAAAGCCT TACCTAGGAA GACGCAGGGT CTAGATAGAA 1440

GCTATAAGGA AGCCACACAC ATAACCCACA TCCCCACACC CCCAACATCC CCCACACTCC 1500

CCACACCCCC CACACCCCCC ACATCCCCAC CATAATTACC CCCACCTCCA AATATCTCAT 1560

AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 1620

AAAAAAA 1627 (2) INFORMATION FOR SEQ ID NO: 14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 65 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Met Gin Ala Glu Val Thr Gly Lys Leu Lys Gin His Phe Asn Asn Ala

1 5 10 15

Leu Pro Lys Gin Asn Ser Thr Thr Thr Gly Val Ser Val He Met Val

20 25 30

Ser Gly Arg Arg Gin Val Trp Pro Asp Cys Gly Ala Gly Leu Glu Val 35 40 45

Phe Glu Leu Gly Ser Arg Glu Leu Pro Ser His Gly Ser Trp Cys Leu 50 55 60 Pro 65

(2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2468 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15: AAATTTTCAT CTCTAAATCA AAAAGTGAAG CAGACCAAAT CCAATGTAAA TATTGGCAAC 60

CTCCGAAAGC TAGGAAACTT TACCAAACCT GAAATGAAAG TTAACTTTCT AAAACCAAAC 120

TTAAAAGTAA ATCTTTGGAA ATCAGATAGT AGTCTTGAAA CTATGGAAAA CACAGGAGTG 180

ATGGATAAGG TTCAGGCAGA GTCTGATGGG GACATGTCTT CAGATAATGA CTCATACCAC 240

TCTGATGAAT TCCTTACAAA TTCTAAGTCT GATGAAGACA GGCAGCTAGC TAACTCATTA 300

GAGAGTGTAG GGCCAATAGA TTACGTTCTT CCTAGTTGTG GTATTATTGC CTCAGCGCCT 360

CGATTGGGCA GTCGGTCCCA GTCTCTTAGC AGCACAGATA GTAGCGTTCA TGCTCCTTCA 420

GAGATTACTG TTGCTCATGG GAGTGGGCTT GGAAAAGGCC AGGAGTCTCC TTTGAAGAAA 480

AGTCCTTCTG CTGGCGACGT ACACATATTG ACTGGCTTTG CCAAGCCTAT GGATATTTAC 540

TGCCACAGAT TTGTGCAAGA TGCACAGAAC AAAGTGACCC ACCTATCAGA GACCAGATCT 600

GTGTCTCAGC AGGCTAGTCA GGAAAGAAAT CAAATGACCA ATCAAGTTTC AAATGAAACC 660

CAATCAGAAT CAACAGAACA GACACCTTCT CGGCCATCGC AATTAGATGT CTCTCTTTCT 720

GCAACAGGCC CACAGTTTTT GTCAGTTGAG CCAGCGCATT CAGTTGCATC TCAAAAAACC 780

CCCACCTCCG CTTCCAGCAT GCTTGAACTT GAGACAGGGC TTCATGTAAC TCCTTCTCCT 840

TCAGAGAGCA GTAGCAGCAG AGCAGTCTCT CCCTTTGCCA AGATTCGAAG TTCCATGGTC 900

CAGGTTGCTA GTATTACCCA AGCTGGATTA ACCCATGGGA TAAACTTTGC AGTGTCAAAA 960

GTTCAGAAGA GTCCTCCAGA ACCTGAAATC ATTAATCAAG TCCAGCAAAA TGAACTTAAA 1020

AAGATGTTTA TACAATGCCA GACACGGATA ATTCAGATTT AGCTTTTAGC CATAAGAATC 1080

CTTCCATGGC TTTTATTTAA AAATATGAAA TTTTCACCTC TTGGGGTATT TTAATTGTAC 1140 TGTCTGAACC CAGGGATCAC AAATTCTGTT CATTGGAAAG GGTTTTAAAC GGAGTCGGAA 1200

CCTGAGTAGA TTTCCAAATT TTACAGCCAG GACTACAGAA GTGCATCATT CTAGAATGTG 1260

TAGACCTGAG TAGCTTATAC ACTACAGAGC ACTTTGCTTA TTTGAAAGTA ATTCAGCAAC 1320

AGGTCACTTT GGGATATAAC CTGAACCTTT TTTTGGAGTG GGGTGGGTAG ACTACAGTAG 1380

ACACAAGGGC TGGACATGCA GATGCTTAGG GGATTAGCGT TTTTCATAAT TTGTTCTGTT 1440

TGTCAGTTCA TTCCTGTGTG TTCTTACCTC TACAAAGTAA ATTACACATT TAGTTTTTAG 1500

TGACTTTAAC ATGTTACTGA AGCATTTGAA TATAAAGCTA TTTTAGTTTT GATGGCTTAA 1560

CTGTTCCCTG AGGAGTTGAG GGTTATTGAC ACTAAAAAAA TGAATTCTCA TTTGATCCTA 1620

ATTTTCCCCG TATTCTACTT GAACACATTA AAAATACTCT GCTGCCTATA CAATGTAAAC 1680

CTAGGAGCAT TAAGACTTGT CACACAGTAA ACCTGATACA TCAGAGGTGA ATACCAGCAC 1740

CTATTAGGTT TCATTTTGCT GTTTTCAGGA ATGTAAGAAC ACCCATATTG GCTACTGGAA 1800

ATTCTAGCAG TCAGTCAGGT TTTAATTTAT TCCAGGAGGG GCATCCTCGA CATCTTATGT 1860

AGATGATCCA CAACTTCAAA ATTTAGTCTG GGCCTAGTGC AGTGGCTCAC ACCTATAATC 1920

CCAACACTTT GGGAGGCCAG GAGTTTTGAG ACCAGCCTGG GAAACATCTG CCTCTACAAA 1980

AAAATACAAA AATTAGCTGG GCATAGTGGT GCATGCCTGT GTTTCTAGCT ACGCAGGAGG 2040

ATTGCTTGAG CCCATGAGAT TGAGGCTGCA GTGAGCTGTG ATCGTGCCAC TGACCTCCAG 2100

CCTGGGGGAC AGAGCAAGAC CGTGTCTCAA AAACAATTTA GTCTGAAACA CAATTGTGCT 2160

GAATCTGTCT GACTATAACT CTGACCACAC AGAACCAGGG CTGCCCCTGT AATCCCCACA 2220

GTAAGAAAGT TGTATGGCAT ATTCCAACAA GTATTGGTTC GTCTGGTGTC TTTAGAGCTT 2280

TACTCTGTTG AAGTGACTGA TTCTCAACTG AACATTATGT CGTTACTTTG ATAAGCATTC 2340

CACTTTTGTT ATTTATTAGT GCTATCTTTT TTTTTTTACG TGTTAAATCT TGTGATTATT 2400

AAAATAAAGT ACCATTGTAA TTTAAAGTGA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA 2460

AAAAAAAA 2468 (2) INFORMATION FOR SEQ ID NO: 16:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 322 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:

Met Lys Val Asn Phe Leu Lys Pro Asn Leu Lys Val Asn Leu Trp Lys

1 5 10 15

Ser Asp Ser Ser Leu Glu Thr Met Glu Asn Thr Gly Val Met Asp Lys 20 25 30

Val Gin Ala Glu Ser Asp Gly Asp Met Ser Ser Asp Asn Asp Ser Tyr 35 40 45

His Ser Asp Glu Phe Leu Thr Asn Ser Lys Ser Asp Glu Asp Arg Gin 50 55 60

Leu Ala Asn Ser Leu Glu Ser Val Gly Pro He Asp Tyr Val Leu Pro 65 70 75 80

Ser Cys Gly He He Ala Ser Ala Pro Arg Leu Gly Ser Arg Ser Gin 85 90 95

Ser Leu Ser Ser Thr Asp Ser Ser Val His Ala Pro Ser Glu He Thr 100 105 110

Val Ala His Gly Sex Gly Leu Gly Lys Gly Gin Glu Ser Pro Leu Lys 115 120 125

Lys Ser Pro Ser Ala Gly Asp Val His He Leu Thr Gly Phe Ala Lys 130 135 140

Pro Met Asp He Tyr Cys His Arg Phe Val Gin Asp Ala Gin Asn Lys 145 150 155 160

Val Thr His Leu Ser Glu Thr Arg Ser Val Ser Gin Gin Ala Ser Gin 165 170 175

Glu Arg Asn Gin Met Thr Asn Gin Val Ser Asn Glu Thr Gin Ser Glu 180 185 190

Ser Thr Glu Gin Thr Pro Ser Arg Pro Ser Gin Leu Asp Val Ser Leu 195 200 205

Ser Ala Thr Gly Pro Gin Phe Leu Ser Val Glu Pro Ala His Ser Val 210 215 220

Ala Ser Gin Lys Thr Pro Thr Ser Ala Ser Ser Met Leu Glu Leu Glu 225 230 235 240

Thr Gly Leu His Val Thr Pro Ser Pro Ser Glu Ser Ser Ser Ser Arg 245 250 255

Ala Val Ser Pro Phe Ala Lys He Arg Ser Ser Met Val Gin Val Ala 260 265 270 Ser He Thr Gin Ala Gly Leu Thr His Gly He Asn Phe Ala Val Ser 275 280 285

Lys Val Gin Lys Ser Pro Pro Glu Pro Glu He He Asn Gin Val Gin 290 295 300

Gin Asn Glu Leu Lys Lys Met Phe He Gin Cys Gin Thr Arg He He 305 310 315 320

Gin He

(2) INFORMATION FOR SEQ ID NO: 17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1044 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: GCGGCCGCGG TGGTCCGGGC CGCCGTGTCT GCTTCCAGAG CCAAATCTGC CGAGGCTGGA 60

ATTGCCGGGG AGGCCCAAAG CAAGAAGCCA GTGTCCAGGC CGGCCACCGC TGCCGCTGCC 120

GCTGCCGGCT CCAGGGAGCC CCGTGTCAAG CAAGGTCCAA AAATTTATAG TTTTAATTCT 180

ACAAATGATT CTAGTGGTCC TGCAAATCTG GATAAATCTA TTTTGAAAGT GGTAATTAAT 240

AACAAACTAG AGCAAAGAAT TATTGGAGTG ATCAATGAGC ATAAAAAGCA AAATAATGAC 300

AAAGGAATGA TTTCTGGAAG ACTTACTGCC AAAAAATTGC AGGATTTATA CATGGCTTTA 360

CAAGCATTTT CATTTAAGAC AAAGGACATT GAAGATGCCA TGACCAATAC ACTCTTATAT 420

GGAGGTGACC TTCATTCTGC CTTGGATTGG CTCTGTTTAA ACCTTTCAGA TGATGCACTT 480

CCTGAAGGAT TCAGTYAGGA ATTWGAAGAG CAGAAACCTA AAAGTAGGCA TAAATTTCAG 540

TCTCCTCAAA TATAAGTTAC TATTTCACCT CCATTGCAAC CTAAAACAAA AACATATGAA 600

GAGGACCCTA AGAGTAAGCC AAAAAAGGAA GAAAAAAATA TGGAAGTAAA TATGAAAGAG 660

TGGATTTTAC GATATGCTGA ACAACAAAAT GAAGAAGAAA AGAATGAGAA TTCTAAAAGT 720

TTAGAAGAGG AGGAAAAATT TGACCCTAAT GAAAGGTACT TACATCTTGC AGCAAAACTG 780

CTGGATGCAA AAGAACAAGC AGCTACCTTT AAACTAGAAA AAAACAAGCA AGGCCAAAAA 840 GAGGCTCAGG AAAAAATAAG GAAATTTCAA AGAGAAATGG AAACTTTAGA AGACCATCCA 900

GTATTTAACC CAGCCATGAA GATTTCACAT CAACAAAATG AAAGGAAAAA GCCTCCTGTA 960

GCCACAGAAG GAGAAAGTGC ATTGAATTTT AATTTATTTG AAAAATCTGC AGCTGCTACT 1020

GAAGAAGAGA AAAAAAAAAA AAAA 1044 (2) INFORMATION FOR SEQ ID NO: 18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 130 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Met Glu Val Asn Met Lys Glu Trp He Leu Arg Tyr Ala Glu Gin Gin 1 5 10 15

Asn Glu Glu Glu Lys Asn Glu Asn Ser Lys Ser Leu Glu Glu Glu Glu 20 25 30

Lys Phe Asp Pro Asn Glu Arg Tyr Leu His Leu Ala Ala Lys Leu Leu 35 40 45

Asp Ala Lys Glu Gin Ala Ala Thr Phe Lys Leu Glu Lys Asn Lys Gin 50 55 60

Gly Gin Lys Glu Ala Gin Glu Lys He Arg Lys Phe Gin Arg Glu Met 65 70 75 80

Glu Thr Leu Glu Asp His Pro Val Phe Asn Pro Ala Met Lys He Ser 85 90 95

His Gin Gin Asn Glu Arg Lys Lys Pro Pro Val Ala Thr Glu Gly Glu 100 105 110

Ser Ala Leu Asn Phe Asn Leu Phe Glu Lys Ser Ala Ala Ala Thr Glu 115 120 125

Glu Glu 130

(2) INFORMATION FOR SEQ ID NO: 19:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 764 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

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

AGGTCTAGAA TTTGTTATTT TAAGTTGAAA TTTGAAGATT GGCTGAGTTG GTTTCATGAT 60

ATTGTTGCCT ACTGATATCT TACACAGAGT ACGAGTACAT CTGGTTGAAA GAGAAAAGGC 120

AGAGAGAAAA GCAGACTTTT TGGAGTTGTT GTTCTCAGGC ACTAACAGAT TCATCCCTTA 180

TATGCATATG AATGGACAAT AAGATTTCTT GGATATTTTC ATGGCAAAGG TGAGGAGAAA 240

TTTTTATTGC TTTTTGAAGA AACATGATTT TTATTACCTT GGGTTATTCC AGGAGATAAT 300

TGATAAATGT TGAGTAGTTT GTTGGTCTTC TTCTTAAAGG AGGATCAAGA GAAGCAGTAC 360

AGCCTGGGTG ACAGAGCGAG ACTCTGTCTC AAATAAAAAA AAAGAAGGAA ACATACTGTA 420

GCCAGACGCC ACCTATAGTC CCAAGTAGCT GGGATTACAG GACAAAACAA TTGAAGAGCT 480

CAGGCAATCT TTAGCAAATG TTGAAAGGAT GAAAGAGAAG GCAAATGTTG AAACGATGAA 540

AGAGAAGGCA GTTGTGAAAA CAGAAAACTT GAAAACTACA TTAGACTCTG CAGAGCAAAA 600

GGCAAGATCA GACAAAGAGA AGACCCAGCA GATGTTAGAT GCTGTCACTT CTGAGCCCCC 660

AACAGCAAAG AGCGCACCTG AAGAAGTATC AGGACAAGAA CAAGAGGTTT TTTCAAAATA 720

GTAAAATTAA AATTAATTTA GTTGAAGTGA TAAAAAAAAA AAAA 764 (2) INFORMATION FOR SEQ ID NO: 20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 70 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:

Met Lys Glu Lys Ala Asn Val Glu Thr Met Lys Glu Lys Ala Val Val 1 5 10 15

Lys Thr Glu Asn Leu Lys Thr Thr Leu Asp Ser Ala Glu Gin Lys Ala 20 25 30

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

Glu Pro Pro Thr Ala Lys Ser Ala Pro Glu Glu Val Ser Gly Gin Glu 50 55 60

Gin Glu Val Phe Ser Lys 65 70

(2) INFORMATION FOR SEQ ID NO: 21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 666 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: GCGAGTCTGT CGATCCCAGG CCAGAGACAA GGCAGACTAA GGTTCATTTG TAAAGAAGCT 60

CCTTCCAGCA CCTCCTCTCT TCTCCTTTTG CCCAAACTCA CCCAGTGAGT GTGAGCATTT 120

AAGAAGCATC CTCTGCCAAG ACCAAAAGGA AAGAAGAAAA AGGGCCAAAA GCCAAAATGA 180

AACTGATGGT ACTTGTTTTC ACCATTGGGC TAACTTTGCT GCTAGGAGTT CAAGCCATGC 240

CTGCAAATCG CCTCTCTTGC TACAGAAAGA TACTAAAAGA TCACAACTGT CACAACCTTC 300

CGGAAGGAGT AGCTGACCTG ACACAGATTG ATGTCAATGT CCAGGATCAT TTCTGGGATG 360

GGAAGGGATG TGAGATGATC TGTTACTGCA ACTTCAGCGA ATTGCTCTGC TGCCCAAAAG 420

ACGTTTTCTT TGGACCAAAG ATCTCTTTCG TGATTCCTTG CAACAATCAA TGAGAATCTT 480

CATGTATTCT GGAGAACACC ATTCCTGATT TCCCACAAAC TGCACTACAT CAGTATAACT 540

GCATTTCTAG TTTCTATATA GTGCAATAGA GCATAGATTC TATAAATTCT TACTTGTCTA 600

AGACAAGTAA ATCTGTGTTA AACAAGTAGT AATAAAAGTT AATTCAATCT AAAAAAAAAA 660

AAAAAA 666 (2) INFORMATION FOR SEQ ID NO: 22:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 98 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22:

Met Lys Leu Met Val Leu Val Phe Thr He Gly Leu Thr Leu Leu Leu 1 5 10 15

Gly Val Gin Ala Met Pro Ala Asn Arg Leu Ser Cys Tyr Arg Lys He 20 25 30

Leu Lys Asp His Asn Cys His Asn Leu Pro Glu Gly Val Ala Asp Leu 35 40 45

Thr Gin He Asp Val Asn Val Gin Asp His Phe Trp Asp Gly Lys Gly 50 55 60

Cys Glu Met He Cys Tyr Cys Asn Phe Ser Glu Leu Leu Cys Cys Pro 65 70 75 80

Lys Asp Val Phe Phe Gly Pro Lys He Ser Phe Val He Pro Cys Asn 85 90 95

Asn Gin

(2) INFORMATION FOR SEQ ID NO: 23:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 28 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: CNAAGCATATA AGTCTCAGGG GCAGGTT 28

(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: CNATTGCTGTG TAACCAATGC AAAACTGA 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: CNCAGAAGAT CCAGGACTAT AAACAGGAT 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: CNGACAAATGT GCAATACAAT CGGTCCTC 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: GNCCATTTGTC TGCTTTGCAT TTTGGATG 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: CNGCTTGCATG TTATTCTCCT GGGAGGCA 29

(2) INFORMATION FOR SEQ ID NO: 29:

(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: 29: CNATCACTCCT GTGTTTTCCA TAGTTTCA 29

(2) INFORMATION FOR SEQ ID NO: 30:

(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: 30: GNTCTTTTGCA TCCAGCAGTT TTGCTGCA 29

(2) INFORMATION FOR SEQ ID NO: 31:

(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:31: CNACATTTGCC TTCTCTTTCA TCCTTTCA 29

(2) INFORMATION FOR SEQ ID NO: 32:

(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: 32: TNAACTCCTAG CAGCAAAGTT AGCCCAAT 29

(2) INFORMATION FOR SEQ ID NO: 33:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 151 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:

Met Ser Gin Asn Met Gly Cys Tyr Phe Leu Phe Leu Ser Trp Phe Pro 1 5 10 15

Tyr Ser His Pro Ser Phe Ser Thr Gin Val Ser Gly Gin Glu Asp Phe 20 25 30

Ser His Gin Leu Tyr Gin Arg Lys Leu Gin Ala Pro Leu Trp Pro Ser 35 40 45

Ser Leu Gly He Thr Asp Cys Cys Gin Tyr Val Thr Ser Cys His Pro 50 55 60

Lys Arg Ser Glu Arg Arg Lys Tyr Gly Arg Asp Phe Leu Leu Arg Phe 65 70 75 80

Arg Phe Cys Ser He Ala Cys Gin Arg Pro Val Gly Leu Val Leu Met 85 90 95

Glu Gly Val Thr Asp Thr Lys Pro Glu Arg Pro Ala Gly Trp Ala Glu 100 105 110

Ser Val Leu Glu Glu Asp Ala Ser Glu Leu Glu Pro Ala Phe Ser Arg 115 120 125

Thr Val Gly Thr He Gin His Cys Leu His Leu Thr Ser Val Tyr Thr 130 135 140

Pro Ser Thr Pro Ser Arg Ala 145 150

(2) INFORMATION FOR SEQ ID NO: 34:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 71 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34:

Met Lys He Gly Leu Ala Cys Gly Gly His Ser Phe Gly Asn Ser Gin 1 5 10 15

Ala Leu Gly Thr Val Trp Arg Gly Leu Cys Arg Asp Gin Gin Ala Trp 20 25 30 Cys Glu Gly Ser Arg Arg Gin Gly Ala Pro Gly Trp Ala Leu Trp Ala 35 40 45

Ala Ala Ser Thr Leu Val Ser Ala Phe Ser Ser Arg Glu Thr Pro Lys 50 55 60

Pro Trp Arg Gly Val Leu Cys 65 70

Claims

What is claimed is:

1. A composition comprising an isolated polynucleotide selected from the group consisting of:

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

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone AA365_1 deposited under accession number ATCC 98296;

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

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

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

(a)-(g) above;

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

3. A host cell transformed with a composition 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 a composition 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 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;

(c) fragments of the amino acid sequence of SEQ ID NO:2; and

(d) the amino acid sequence encoded by the cDNA insert of clone AA365_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

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

10. The composition of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2 from amino acid 76 to amino acid 109.

11. The composition of claim 8, further comprising 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. 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 245 to nucleotide 421;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 from nucleotide 413 to nucleotide 421;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone BL67_2 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296;

(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;

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

15. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:

(c) fragments of the amino acid sequence of SEQ ID NO:4; and

(d) the amino acid sequence encoded by the cDNA insert of clone BL67_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

16. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:3 or SEQ ID NO:5.

17. A composition comprising 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 137 to nucleotide 2146;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:9 from nucleotide 218 to nucleotide 2146;

(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CG99_2 deposited under accession number ATCC 98296;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity; (k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

18. 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:10;

(b) the amino acid sequence of SEQ ID NO:10 from amino acid 50 to amino acid 87;

(c) fragments of the amino acid sequence of SEQ ID NO: 10; and

(d) the amino acid sequence encoded by the cDNA insert of clone CG99_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

19. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:9.

20. A composition comprising an isolated polynucleotide selected from the group consisting of:

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

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:ll from nucleotide 123 to nucleotide 1247;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 from nucleotide 1 to nucleotide 419;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone CO618_l deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CO618_l deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296;

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

(a)-(g) above;

21. 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: 12;

(b) the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 99;

(c) fragments of the amino acid sequence of SEQ ID NO:12; and

(d) the amino acid sequence encoded by the cDNA insert of clone CO618_l deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

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

23. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:13;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:14 having biological activity;

24. 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:14;

(b) the amino acid sequence of SEQ ID NO:14 from amino acid 1 to amino acid 31;

(c) fragments of the amino acid sequence of SEQ ID NO:14; and

(d) the amino acid sequence encoded by the cDNA insert of clone CO629_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

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

26. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:15;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 15 from nucleotide 1 to nucleotide 387;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16;

(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 16 having biological activity;

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

(a)-(g) above;

27. 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:16;

(b) the amino acid sequence of SEQ ID NO:16 from amino acid 1 to amino acid 98; (c) fragments of the amino acid sequence of SEQ ID NO:16; and

(d) the amino acid sequence encoded by the cDNA insert of clone CT645_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

28. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:15.

29. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17 from nucleotide 640 to nucleotide 1029;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:17 from nucleotide 611 to nucleotide 814;

(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CW383_1 deposited under accession number ATCC 98296;

30. 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: 18;

(b) the amino acid sequence of SEQ ID NO: 18 from amino acid 1 to amino acid 63;

(c) fragments of the amino acid sequence of SEQ ID NO: 18; and

(d) the amino acid sequence encoded by the cDNA insert of clone CW383_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

31. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:17.

32. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:19;

(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DN167_1 deposited under accession number ATCC 98296;

(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296;

33. 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:20;

(b) the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 65;

(c) fragments of the amino acid sequence of SEQ ID NO:20; and

(d) the amino acid sequence encoded by the cDNA insert of clone DN167_1 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

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

35. A composition comprising an isolated polynucleotide selected from the group consisting of:

(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:21;

(b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:21 from nucleotide 177 to nucleotide 470;

(c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:21 from nucleotide 237 to nucleotide 470;

(d) a polynucleotide comprising the nucleotide sequence of the full- length protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296;

(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296; (f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DN711_2 deposited under accession number ATCC 98296;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296;

(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:22;

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

(a)-(g) above;

36. 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:22;

(b) the amino acid sequence of SEQ ID NO:22 from amino acid 1 to amino acid 94;

(c) fragments of the amino acid sequence of SEQ ID NO.22; and

(d) the amino acid sequence encoded by the cDNA insert of clone DN711_2 deposited under accession number ATCC 98296; the protein being substantially free from other mammalian proteins.

37. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:21.