CA2294569A1

CA2294569A1 - Secreted proteins and polynucleotides encoding them

Info

Publication number: CA2294569A1
Application number: CA002294569A
Authority: CA
Inventors: Kenneth Jacobs; John M. Mccoy; Edward R. Lavallie; Lisa A. Racie; Maurice Treacy; Vikki Spaulding; Michael J. Agostino; Steven H. Howes; Kim Fechtel
Original assignee: Individual
Current assignee: Genetics Institute LLC
Priority date: 1997-06-19
Filing date: 1998-06-18
Publication date: 1998-12-23
Also published as: EP1003770A1; AU7970498A; WO1998057976A1; EP1003770A4; JP2002513294A

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 application Ser. No. 60/XXX,XXX
(converted to a provisional application from non-provisional application Ser.
No.
08/878,715), filed June 19, 1997, which is incorporated by reference herein.
FIELD OF THE INVENTION
The present invention provides novel polynucleotides and proteins encoded by such polynucleotides, along with therapeutic, diagnostic and research utilities for these polynucleotides and proteins.
BACKGROUND OF THE INVENTION
Technology aimed at the discovery of protein factors (including e.g., cytokines, such as lymphokines, interferons, CSFs and interleukins) has matured rapidly over the past decade. The now routine hybridization cloning and expression cloning techniques clone novel polynucleotides "directly" in the sense that they rely on information directly related to the discovered protein (i.e., partial DNA/amino acid sequence of the protein in the case of hybridization cloning; activity of the protein in the case of expression cloning). More recent "indirect" cloning techniques such as signal sequence cloning, which isolates DNA sequences based on the presence of a now well-recognized secretory leader sequence motif, as well as various PCR-based or low stringency hybridization cloning techniques, have advanced the state of the art by making available large numbers of DNA/amino acid sequences for proteins that are known to have biological activity by virtue of their secreted nature in the case of leader sequence cloning, or by virtue of the cell or tissue source in the case of PCR-based techniques. It is to these proteins and the polynucleotides encoding them that the present invention is directed.
SUBSTITUTE SHEET (RULE 26) 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
N0:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 185 to nucleotide 1600;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 1403 to nucleotide 1600;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:1 from nucleotide 1 to nucleotide 850;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone dol5 4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone dol5 4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone dol5_4 deposited under accession number ATCC 98468;

2 0 (h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone dol5 4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2;
(j) a polynucleotide encoding a protein comprising a fragment of the 2 5 amino acid sequence of SEQ ID N0:2 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:2;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(1) a polynucleotide which encodes a species homologue of the protein 3 0 of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-{j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO:1 from nucleotide 185 to nucleotide 1600; the nucleotide sequence of SEQ ID
N0:1 from nucleotide 1403 to nucleotide 1600; the nucleotide sequence of SEQ ID
N0:1 from nucleotide 1 to nucleotide 850; the nucleotide sequence of the full-length protein coding sequence of clone dol5_4 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone dol5 4 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone dol5_4 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2 from amino acid 1 to amino acid 222. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:2, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment comprising the amino acid sequence from amino acid 231 to amino acid 240 of SEQ ID N0:2.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:1.
In other embodiments, the present invention provides a composition comprising 2 0 a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:2;
(b) the amino acid sequence of SEQ ID N0:2 from amino acid 1 to amino acid 222;
2 5 (c) fragments of the amino acid sequence of SEQ ID N0:2 comprising eight consecutive amino acids of SEQ ID N0:2; and (d) the amino acid sequence encoded by the cDNA insert of clone dol5 4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such 3 0 protein comprises the amino acid sequence of SEQ ID N0:2 or the amino acid sequence of SEQ ID N0:2 from amino acid 1 to amino acid 222. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID

N0:2, or a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:2having biological activity, the fragment comprising the amino acid sequence from amino acid 231 to amino acid 240 of SEQ ID N0:2.
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
N0:3;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 47 to nucleotide 2065;
(c} a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 1086 to nucleotide 1848;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone dx290_1 deposited under accession number ATCC 98468;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468;
2 0 (g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone dx290_1 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:4;
(i) a polynucleotide encoding a protein comprising a fragment of the 2 5 amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:4;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein 3 0 of (h) or (i) above ; and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:3 from nucleotide 47 to nucleotide 2065; the nucleotide sequence of SEQ ID
N0:3 from nucleotide 1086 to nucleotide 1848; the nucleotide sequence of the full-length protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468;
or the nucleotide sequence of a mature protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone dx290_1 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:4 from amino acid 312 to amino acid 600. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:4 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:4, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment comprising the amino acid sequence from amino acid 331 to amino acid 340 of SEQ ID N0:4.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:3.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 2 0 consisting of:
(a) the amino acid sequence of SEQ ID N0:4;
(b) the amino acid sequence of SEQ ID N0:4 from amino acid 312 to amino acid 600;
(c) fragments of the amino acid sequence of SEQ ID N0:4 comprising 2 5 eight consecutive amino acids of SEQ ID N0:4; and (d) the amino acid sequence encoded by the cDNA insert of clone dx290_1 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:4 or the amino acid sequence 3 0 of SEQ ID N0:4 from amino acid 312 to amino acid 600. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:4, or a protein comprising a fragment of the amino acid sequence of SEQ ID N0:4having biological activity, the fragment comprising the amino acid sequence from amino acid 331 to amino acid 340 of SEQ ID N0:4.
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
N0:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 107 to nucleotide 724;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 218 to nucleotide 724;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 536 to nucleotide 866;
{e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ek390 4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ek390 4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ek390_4 deposited under accession number 2 0 ATCC 98468;
{h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ek390 4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:6;
2 5 (j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:6;
{k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
3 0 (1) a polynucleotide which encodes a species homologue of the protein of {i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:5 from nucleotide 107 to nucleotide 724; the nucleotide sequence of SEQ ID
N0:5 from nucleotide 218 to nucleotide 724; the nucleotide sequence of SEQ ID N0:5 from nucleotide 536 to nucleotide 866; the nucleotide sequence of the full-length protein coding sequence of clone ek390 4 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone ek390_4 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone ek390 4 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:6 from amino acid 6 to amino acid 92.
In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:6 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:6, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment comprising the amino acid sequence from amino acid 97 to amino acid 106 of SEQ ID N0:6.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
2 0 ID N0:5.
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 N0:6;
2 5 (b) the amino acid sequence of SEQ ID N0:6 from amino acid 6 to amino acid 92;
(c) fragments of the amino acid sequence of SEQ ID N0:6 comprising eight consecutive amino acids of SEQ ID N0:6; and (d) the amino acid sequence encoded by the cDNA insert of clone 3 0 ek390 4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:6 or the amino acid sequence of SEQ ID N0:6 from amino acid 6 to amino acid 92. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:6 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID
N0:6, or a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:6having biological activity, the fragment comprising the amino acid sequence from amino acid 97 to amino acid 106 of SEQ ID N0:6.
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
N0:7;
1 p (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 31 to nucleotide 1230;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 289 to nucleotide 1230;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 344 to nucleotide 1119;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone er471 7 deposited under accession number ATCC 98468;
{f) a polynucleotide encoding the full-length protein encoded by the 2 0 cDNA _insert of clone er471 7 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone er471 7 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
2 5 insert of clone er471 7 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment 3 0 comprising eight consecutive amino acids of SEQ ID N0:8;
(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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:7 from nucleotide 31 to nucleotide 1230; the nucleotide sequence of SEQ ID
N0:7 from nucleotide 289 to nucleotide 1230; the nucleotide sequence of SEQ ID N0:7 from nucleotide 344 to nucleotide 1119; the nucleotide sequence of the full-length protein coding sequence of clone er471 7 deposited under accession number ATCC 98468;
or the nucleotide sequence of a mature protein coding sequence of clone er471 7 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone er471 7 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8 from amino acid 111 to amino acid 363. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:8, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment comprising the amino acid sequence 2 0 from amino acid 195 to amino acid 204 of SEQ ID N0:8.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:7.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 2 5 consisting of:
(a) the amino acid sequence of SEQ ID N0:8;
(b) the amino acid sequence of SEQ ID N0:8 from amino acid 111 to amino acid 363;
(c) fragments of the amino acid sequence of SEQ ID N0:8 comprising 3 0 eight consecutive amino acids of SEQ ID N0:8; and (d) the amino acid sequence encoded by the cDNA insert of clone er471 7 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:B or the amino acid sequence of SEQ ID N0:8 from amino acid 111 to amino acid 363. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:8, or a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8having biological activity, the fragment comprising the amino acid sequence from amino acid 195 to amino acid 204 of SEQ ID N0:8.
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
N0:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 62 to nucleotide 322;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 571 to nucleotide 878;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone fs40 3 deposited under accession number ATCC 98468;
{e) a polynucleotide encoding the full-length protein encoded by the 2 0 cDNA insert of clone fs40_3 deposited under accession number ATCC 98468;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone fs40 3 deposited under accession number ATCC
98468;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
2 5 insert of clone fs40_3 deposited under accession number ATCC 98468;
{h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment 3 0 comprising eight consecutive amino acids of SEQ ID N0:10;
(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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:9 from nucleotide 62 to nucleotide 322; the nucleotide sequence of SEQ ID
N0:9 from nucleotide 571 to nucleotide 878; the nucleotide sequence of the full-length protein coding sequence of clone fs40 3 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone fs40_3 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone fs40 3 deposited under accession number ATCC 98468. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:10, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 38 to amino acid 47 of SEQ
ID NO:10.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:9.
2 0 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 N0:10;
(b) fragments of the amino acid sequence of SEQ ID NO:10 comprising 2 5 eight consecutive amino acids of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone fs40 3 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:10. In further preferred 3 0 embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:10, or a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity, the fragment comprising the amino acid sequence from amino acid 38 to amino acid 47 of SEQ ID N0:10.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11 from nucleotide 43 to nucleotide 1671;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11 from nucleotide 112 to nucleotide 1671;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:11 from nucleotide 224 to nucleotide 679;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ga63_6 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA _insert of clone ga63_6 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ga63_6 deposited under accession number ATCC
2 0 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ga63 6 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12;
2 5 (j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:12;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
3 0 (1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in {a)-(j).

Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:11 from nucleotide 43 to nucleotide 1671; the nucleotide sequence of SEQ ID
N0:11 from nucleotide 112 to nucleotide 1671; the nucleotide sequence of SEQ ID
N0:11 from nucleotide 224 to nucleotide 679; the nucleotide sequence of the full-length protein coding sequence of clone ga63 6 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone ga63_6 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone ga63_6 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12 from amino acid 62 to amino acid 212. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:12, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:12 having biological activity, the fragment comprising the amino acid sequence from amino acid 266 to amino acid 275 of SEQ ID N0:12.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
2 0 ID NO:11.
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 N0:12;
2 5 {b) the amino acid sequence of SEQ ID N0:12 from amino acid 62 to amino acid 212;
(c) fragments of the amino acid sequence of SEQ ID N0:12 comprising eight consecutive amino acids of SEQ ID N0:12; and (d) the amino acid sequence encoded by the cDNA insert of clone 3 0 ga63_6 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:12 or the amino acid sequence of SEQ ID N0:12 from amino acid 62 to amino acid 212. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:12, or a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12having biological activity, the fragment comprising the amino acid sequence from amino acid 266 to amino acid 275 of SEQ ID N0:12.
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
N0:13;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 17 to nucleotide 523;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 77 to nucleotide 523;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:13 from nucleotide 1 to nucleotide 392;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone gm335 4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the 2 0 cDNA insert of clone gm335 4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone gm335 4 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
2 5 insert of clone gm335 4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:14;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment 3 0 comprising eight consecutive amino acids of SEQ ID N0:14;
(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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
. N0:13 from nucleotide 17 to nucleotide 523; the nucleotide sequence of SEQ
ID N0:13 from nucleotide 77 to nucleotide 523; the nucleotide sequence of SEQ ID N0:13 from nucleotide 1 to nucleotide 392; the nucleotide sequence of the full-length protein coding sequence of clone gm335 4 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone gm335_4 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone gm335 4 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:14 from amino acid 1 to amino acid 125. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:14 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:14, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment comprising the amino acid 2 0 sequence from amino acid 79 to amino acid 88 of SEQ ID N0:14.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:13.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group 2 5 consisting of:
(a) the amino acid sequence of SEQ ID N0:14;
(b) the amino acid sequence of SEQ ID N0:14 from amino acid 1 to amino acid 125;
{c) fragments of the amino acid sequence of SEQ ID N0:14 comprising 3 0 eight consecutive amino acids of SEQ ID N0:14; and (d) the amino acid sequence encoded by the cDNA insert of clone gm335_4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:14 or the amino acid sequence of SEQ ID N0:14 from amino acid 1 to amino acid 125. 1n further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID
N0:14, or a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:14having biological activity, the fragment comprising the amino acid sequence from amino acid 79 to amino acid 88 of SEQ ID N0:14.
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
N0:15;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:I5 from nucleotide 2 to nucleotide 991;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 62 to nucleotide 991;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 2 to nucleotide 504;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone hy370 9 deposited under accession 2 0 number ATCC 98468;
{f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone hy370_9 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone hy370_9 deposited under accession number 2 5 ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone hy370 9 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:16;
3 0 (j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:16;
(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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:15 from nucleotide 2 to nucleotide 991; the nucleotide sequence of SEQ ID
N0:15 from nucleotide 62 to nucleotide 991; the nucleotide sequence of SEQ ID N0:15 from nucleotide 2 to nucleotide 504; the nucleotide sequence of the full-length protein coding sequence of clone hy370_9 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone hy370 9 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone hy370 9 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:16 from amino acid 1 to amino acid 167. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:16 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:16, or a 2 0 polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment comprising the amino acid sequence from amino acid 160 to amino acid 169 of SEQ ID N0:16.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:15.
2 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 N0:16;
(b) the amino acid sequence of SEQ ID N0:16 from amino acid 1 to 3 0 amino acid 167;
(c) fragments of the amino acid sequence of SEQ ID N0:16 comprising eight consecutive amino acids of SEQ ID N0:16; and (d) the amino acid sequence encoded by the cDNA insert of clone hy370_9 deposited under accession number ATCC 98468;

the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:16 or the amino acid sequence of SEQ ID N0:16 from amino acid 1 to amino acid 167. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID
N0:16, or a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:16having biological activity, the fragment comprising the amino acid sequence from amino acid 160 to amino acid 169 of SEQ ID N0:16.
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
N0:17;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 77 to nucleotide 616;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 164 to nucleotide 616;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:17 from nucleotide 1 to nucleotide 415;
2 0 (e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ie47_4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ie47 4 deposited under accession number ATCC 98468;
2 5 (g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ie47 4 deposited under accession number ATCC
98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ie47 4 deposited under accession number ATCC 98468;
3 0 (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:18;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:18;

{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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:17 from nucleotide 77 to nucleotide 616; the nucleotide sequence of SEQ ID
N0:17 from nucleotide 164 to nucleotide 616; the nucleotide sequence of SEQ ID N0:17 from nucleotide 1 to nucleotide 415; the nucleotide sequence of the full-length protein coding sequence of clone ie47_4 deposited under accession number ATCC 98468; or the nucleotide sequence of a mature protein coding sequence of clone ie47 4 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA insert of clone ie47_4 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:18 from amino acid 1 to amino acid 113. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological 2 0 activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:18, or a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment comprising the amino acid sequence from amino acid 85 to amino acid 94 of SEQ ID N0:18.
2 5 Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:17.
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:
3 0 (a) the amino acid sequence of SEQ ID N0:18;
(b) the amino acid sequence of SEQ ID N0:18 from amino acid 1 to amino acid 113;
{c) fragments of the amino acid sequence of SEQ ID N0:18 comprising eight consecutive amino acids of SEQ ID N0:18; and (d) the amino acid sequence encoded by the cDNA insert of clone ie47 4 deposited under accession number ATCC 98468;
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 N0:18 from amino acid 2 to amino acid 113. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID
N0:18, or a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:18having biological activity, the fragment comprising the amino acid sequence from amino acid 85 to amino acid 94 of SEQ ID N0:18.
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
N0:19;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 564 to nucleotide 2813;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 705 to nucleotide 2813;
2 0 (d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 793 to nucleotide 1628;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone s195_10 deposited under accession number ATCC 98468;
2 5 (f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone s195_10 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone s195_10 deposited under accession number ATCC 98468;
3 0 (h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone s195_10 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:20;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID N0:20;
(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 that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:19 from nucleotide 564 to nucleotide 2813; the nucleotide sequence of SEQ
ID N0:19 from nucleotide 705 to nucleotide 2813; the nucleotide sequence of SEQ ID
N0:19 from nucleotide 793 to nucleotide 1628; the nucleotide sequence of the full-length protein coding sequence of clone s195_10 deposited under accession number ATCC 98468;
or the 1 S nucleotide sequence of a mature protein coding sequence of clone s195_10 deposited under accession number ATCC 98468. In other preferred embodiments, the polynucleotide encodes the full-length or a mature protein encoded by the cDNA
insert of clone s195_10 deposited under accession number ATCC 98468. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein 2 0 comprising the amino acid sequence of SEQ ID N0:20 from amino acid 78 to amino acid 355. In further preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID
N0:20 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:20, or a 2 5 polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment comprising the amino acid sequence from amino acid 370 to amino acid 379 of SEQ ID N0:20.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:19.
3 0 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 N0:20;

(b) the amino acid sequence of SEQ ID N0:20 from amino acid 78 to amino acid 355;
(c) fragments of the amino acid sequence of SEQ ID N0:20 comprising eight consecutive amino acids of SEQ ID N0:20; and (d) the amino acid sequence encoded by the cDNA insert of clone s195_10 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:20 or the amino acid sequence of SEQ ID N0:20 from amino acid 78 to amino acid 355. In further preferred embodiments, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20 having biological activity, the fragment preferably comprising eight (more preferably twenty, most preferably thirty) consecutive amino acids of SEQ ID N0:20, or a protein comprising a fragment of the amino acid sequence of SEQ ID N0:20having biological activity, the fragment comprising the amino acid sequence from amino acid 370 to amino acid 379 of SEQ ID N0:20.
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 2 0 modified expression of the genes) 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 2 5 (b) purifying the protein from the culture.
The protein produced according to such methods is also provided by the present invention.
Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier. Compositions comprising an antibody which 3 0 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 lA and 1B 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 forms) 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 2 0 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.
2 5 Clone "dol5 4"
A polynucleotide of the present invention has been identified as clone "dol5 4".
dol5 4 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 3 0 analysis of the amino acid sequence of the encoded protein. dol5_4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "dol5 4 protein").
The nucleotide sequence of dol5 4 as presently determined is reported in SEQ
ID
N0:1. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the dol5 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:2. Amino acids 394 to 406 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 407, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone dol5 4 should be approximately 1900 bp.
The nucleotide sequence disclosed herein for dol5 4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. dol5_4 demonstrated at least some similarity with sequences identified as AA 113909 (zm80f 12.r1 Stratagene neuroepithelium (#937231 ) Homo sapiens cDNA clone 531983 5'), AA189888 (mu55h06.r1 Soares mouse lymph node NbMLN Mus musculus cDNA clone 643355 5'), and U52052 (Human S6 A-8 mRNA expressed in chromosome 6-suppressed melanoma cells). Based upon sequence similarity, dol5 proteins and each similar protein or peptide may share at least some activity.
Clone "dx290 1"
A polynucleotide of the present invention has been identified as clone "dx290_1".
dx290_1 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 2 0 identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. dx290_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "dx290_1 protein").
The nucleotide sequence of dx290_1 as presently determined is reported in SEQ
2 5 ID N0:3. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the dx290_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:4.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone dx290_1 should be approximately 2300 bp.
3 0 The nucleotide sequence disclosed herein for dx290_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. dx290_1 demonstrated at least some similarity with the sequence identified as AA064383 (m147h02.rI Stratagene mouse testis (#937308) Mus musculus cDNA clone 515187 5'). Based upon sequence similarity, dx290_1 proteins and each similar protein or peptide may share at least some activity.
Clone "ek390 4"
A polynucleotide of the present invention has been identified as clone "ek390 4".
ek390_4 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. ek390 4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ek390 4 protein"}.
The nucleotide sequence of ek390 4 as presently determined is reported in SEQ
ID N0:5. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the ek390 4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:6. Amino acids 25 to 37 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 38, or are a transmembrane domain.
The EcoIZI/NotI restriction fragment obtainable from the deposit containing clone ek390_4 should be approximately 1000 bp.
2 0 The nucleotide sequence disclosed herein for ek390 4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. ek390_4 demonstrated at least some similarity with sequences identified as AA075783 {zm89h02.r1 Stratagene ovarian cancer (#937219) Homo sapiens cDNA clone 545139 5'), AA427538 (zw32g04.r1 Soares ovary tumor NbHOT Homo 2 5 sapiens cDNA clone 771030 5'), AA427539 (zw32g04.s 1 Soares ovary tumor NbHOT
Homo Sapiens cDNA clone 771030 3'), AA453353 (zx47a06.r1 Soares testis NHT
Homo sapiens cDNA clone 795346 5'), C20637 (HUMGS0004639, Human Gene Signature, 3'-directed cDNA sequence), R?4326 (y101c07.s1 Homo sapiens cDNA clone 156972 3'), 874420 (ylO1c07.r1 Homo sapiens cDNA clone 156972 5'), T22914 (Human gene 3 0 signature), U41197 (Human [TTTC] 10 short tandem repeat polymorphism UM65, D 1751340), and X58237 (Human mRNA for anti-lectin antibody epitope (clone p36/8-6)).
Based upon sequence similarity, ek390 4 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts a potential transmembrane domain within the ek390 4 protein sequence centered around amino acid 160 of SEQ ID N0:6. The nucleotide sequence of ek390 4 indicates that it may contain GGGA repeat sequences.
Clone "er471 7"
A polynucleotide of the present invention has been identified as clone "er471 7".
er471 7 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins {see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. er471 7 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "er471 7 protein").
The nucleotide sequence of er471 7 as presently determined is reported in SEQ
ID
N0:7. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the er471 7 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:8. Amino acids 74 to 86 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 87, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone 2 0 er471 7 should be approximately 2250 bp.
The nucleotide sequence disclosed herein for er471 7 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. er471 7 demonstrated at least some similarity with sequences identified as AA039137 (mi98h06.r1 Soares mouse embryo NbME13.5 14.5 Mus 2 5 musculus cDNA clone 474683 5'), AA066962 (mm38g05.r1 Stratagene mouse melanoma (#937312) Mus musculus cDNA clone 523832 5'), AA189170 (zq47hO5.s1 Stratagene hNT neuron (#937233) Homo sapiens cDNA clone 632889 3'), AA609188 (af12c10.s1 Soares testis NHT Homo sapiens cDNA clone 1031442 3'), and W07704 (zb02e02.r1 Soares fetal lung NbHLI9W Homo sapiens cDNA clone 300890 5' similar to 3 0 SW:YN66_YEAST P40164 HYPOTHETICAL 98.1 KD PROTEIN IN SPX19-GCR2 INTERGENIC REGION). The predicted amino acid sequence disclosed herein for er471 7 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted er471 7 protein demonstrated at least some similarity to sequences identified as AF016448 (Cosmid F41E6 [Caenorhabditis elegans]) and L08407 (collagen type XVII [Mus musculus]). Based upon sequence similarity, er471 7 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts three potential transmembrane domains within the er471 7 protein sequence, centered around amino acids 40, 80, and 110 of SEQ ID N0:8, respectively.
Clone "fs40 3"
A polynucleotide of the present invention has been identified as clone "fs40 3".
fs40 3 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. fs40 3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "fs40_3 protein").
The nucleotide sequence of fs40_3 as presently determined is reported in SEQ
ID
N0:9. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the fs40 3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:10.
2 0 The EcoRI/NotI restriction fragment obtainable from the deposit containing clone fs40_3 should be approximately 1000 bp.
The nucleotide sequence disclosed herein for fs40_3 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. fs40_3 demonstrated at least some similarity with sequences 2 5 identified as AA411142 (zt37gOl .rl Soares ovary tumor NbHOT Homo sapiens cDNA
clone 724560 5'), AA412527 (zu 12a03.s 1 Soares testis NHT Homo sapiens cDNA
clone 731596 3'), AA565855 (nj32d09.s1 NCI CGAP_AAl Homo sapiens cDNA clone IMAGE:994193), H17042 (ym39f12.s1 Homo sapiens cDNA clone 50584 3'), and (EST57284 Homo sapiens cDNA 3' end similar to None). Based upon sequence similarity, 3 0 fs40_3 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts a potential transmembrane domain within the fs40_3 protein sequence at the C-terminus of SEQ ID N0:10.

WO 98/57976 PCTlUS98/12516 Clone "ga63 6"
A polynucleotide of the present invention has been identified as clone "ga63_6".
ga63_6 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. ga63_6 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ga63_6 protein") The nucleotide sequence of ga63_6 as presently determined is reported in SEQ
ID
N0:11. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the ga63_6 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:12. Amino acids 11 to 23 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 24, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone ga63_6 should be approximately 2300 bp.
The nucleotide sequence disclosed herein for ga63_6 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. ga63_6 demonstrated at least some similarity with sequences identified as AA405433 (zu13h10.r1 Soares testis NHT Homo sapiens cDNA clone 731779 5'similar to TR 6474970 6474970 SP32 PRECURSOR), AA406076 (zu67c02.s1 Soares testis NHT Homo Sapiens cDNA clone 743042 3' similar to TR:G475021 SP32 PRECURSOR), AA424694 (zu13h10.sI Soares testis NHT Homo sapiens cDNA
clone 731779 3' similar to TR 6475021 6475021 SP32 PRECURSOR; contains element 2 5 TAR 1 repetitive element), D 16200 (Pig mRNA for sp32, partial sequence), (Guinea pig mRNA for sp32, complete cds), and D 17573 (Mouse mRNA for proacrosin-binding protein (sp32), complete cds). The predicted amino acid sequence disclosed herein for ga63_6 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted ga63_6 protein 3 0 demonstrated at least some similarity to sequences identified as D16200 (sp32 precursor [Sus scrofa]), and D17574 (alternative splicing product for proacrosin-binding protein (sp32) [Mus musculus]). The sp32 protein is found in the acrosomal vescicle of sperm, which is involved in egg-sperm fusion in fertilization. This protein is initially synthesized as a 61-kDa precursor protein with a putative signal peptide at the amino terminus. The carboxyl-terminal half of the precursor molecule corresponds to the mature sp32 protein.
Thus, sp32 is produced by post-translational modification of the precursor.
The binding of sp32 to proacrosin may be involved in packaging the acrosin zymogen into the acrosomal matrix. (Baba et al., 1994, J. Biol. Chem. 269 (13): 10133-10140, which is incorporated by reference herein). Based upon sequence similarity, ga63_6 proteins and each similar protein or peptide may share at least some activity.
Clone "gm335 4"
A polynucleotide of the present invention has been identified as clone "gm335 4".
gm335 4 was isolated from a human adult uterus 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. gm335 4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "gm335 4 protein") The nucleotide sequence of gm335 4 as presently determined is reported in SEQ
ID N0:13. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the gm335 4 protein corresponding to the foregoing 2 0 nucleotide sequence is reported in SEQ ID N0:14. Amino acids 8 to 20 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 21, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone gm335_4 should be approximately 800 bp.
2 5 The nucleotide sequence disclosed herein for gm335_4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. gm335 4 demonstrated at least some similarity with sequences identified as AAOSS367 (zf20bOS.r1 Soares fetal heart NbHHI9W Homo sapiens cDNA
clone 377457 S'), AC002389 (Human DNA from chromosome 19 specific cosmid 828461, 3 0 genomic sequence, complete sequence), W08S22 (mb46h10.r1 Soares mouse p3NMF19.S
Mus musculus cDNA clone 332S1S S'), and X93916 (S.scrofa mRNA (clone VIB11;
expressed sequence tag)). Based upon sequence similarity, gm335_4 proteins and each similar protein or peptide may share at least some activity.

Clone "hv370 9"
A polynucleotide of the present invention has been identified as clone "hy370_9".
hy370 9 was isolated from a human adult trachea 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. hy370_9 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "hy370 9 protein") The nucleotide sequence of hy370 9 as presently determined is reported in SEQ
ID N0:15. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the hy370 9 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:16. Amino acids 8 to 20 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 21, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone hy370 9 should be approximately 1200 bp.
The nucleotide sequence disclosed herein for hy370 9 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. hy370_9 demonstrated at least some similarity with the sequence identified as AA763313 (vv89h07.r1 Stratagene mouse skin (#937313) Mus musculus cDNA clone 1229629 5'). Based upon sequence similarity, hy370_9 proteins and each similar protein or peptide may share at least some activity. The TopPredII
computer program predicts an additional potential transmembrane domain within the hy370_9 protein sequence centered around amino acid 140 of SEQ ID N0:16.
Clone "ie47 4"
A polynucleotide of the present invention has been identified as clone "ie47_4".
ie47 4 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 3 0 identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. ie47 4 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "ie47_4 protein") The nucleotide sequence of ie47_4 as presently determined is reported in SEQ
ID
N0:27. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the ie47_4 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:18. Amino acids 17 to 29 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 30, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone ie47_4 should be approximately 2300 bp.
The nucleotide sequence disclosed herein for ie47_4 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. ie47_4 demonstrated at least some similarity with sequences identified as AA071953 (mf17h08.r1 Life Tech mouse brain Mus musculus cDNA
clone 405375 5' similar to TR 6304421 6304421 SILENCER ELEMENT), AA207250 (zq82dO5.s1 Stratagene hNT neuron (#937233) Homo Sapiens cDNA clone 648105 3' similar to TR 6304421 6304421 SILENCER ELEMENT), L14938 (Chicken SCG10 protein mRNA, complete cds), L20260 (Mouse SCG10 gene sequence), 849053 (yg58cO5.s1 Homo Sapiens cDNA clone 37017 3'), S82024 (SCG10 neuron-specific growth-associated protein/stathmin homolog [human, embryo, mRNA]), T25428 (Human gene signature HUMGS07594, T25428 standard; cDNA to mRNA), W54204 (md04al2.rl 2 0 Soares mouse embryo NbME13.5 14.5 Mus musculus cDNA clone 367390 5' similar to SW:SCGB XENLA Q09002 SCG10 PROTEIN HOMOLOG A), X71433 (X. laevis SCG 10 mRNA), and 299916 (Human DNA sequence *** SEQUENCING IN PROGRESS
* * * from clone 221 G9; HTGS phase 1 ). The predicted amino acid sequence disclosed herein for ie47_4 was searched against the GenPept and GeneSeq amino acid sequence 2 5 databases using the BLASTX search protocol. The predicted ie47 4 protein demonstrated at least some similarity to sequences identified as L14938 (SCG 10 protein [Gallus gallus]) and S82024 (SCG10 neuron-specific growth-associated protein/stathmin homolog [human, embryo, Peptide] [Homo Sapiens]). SCG10 protein is considered to be a membrane-bound protein present in neural growth cones and developing neurons (Maucuer et al., 1993, J.
3 0 Biol. Chem. 268: 16420-16429; Stein et al.,1988, Neuron 1:463-476; which are incorporated by reference herein). Based upon sequence similarity, ie47_4 proteins and each similar protein or peptide may share at least some activity.

WO 98/57976 PCT/iJS98/12516 Clone "s195 10"
A polynucleotide of the present invention has been identified as clone "s195_10".
s195_10 was isolated from a human adult neural tissue 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. s195_10 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "s195_10 protein").
The nucleotide sequence of s195_10 as presently determined is reported in SEQ
ID
N0:19. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the s195_10 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:20. Amino acids 35 to 47 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 48, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone s195 10 should be approximately 3500 bp.
The nucleotide sequence disclosed herein for s195_10 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. s195_10 demonstrated at least some similarity with sequences 2 0 identified as AA 113800 (zn65bO5.s 1 Stratagene HeLa cell s3 937216 Homo sapiens cDNA clone 563025 3' similar to TR:G600018 6600018 SSM4P), AA114062 (zn65bO5.rl Stratagene HeLa cell s3 937216 Homo sapiens cDNA clone 563025 5'), AA280316 (zt10f06.s1 Soares NbHTGBC Homo Sapiens cDNA clone 712739 3'), AF009301 (Homo sapiens TEB4 protein mRNA, complete cds), N70344 (za60f 10. s 1 Homo Sapiens cDNA
clone 296971 3'), 860474 (yh13g07.r1 Homo sapiens cDNA clone 43058 5'), and (standard; cDNA to mRNA; 148 BP, Human gene signature HUMGS08505). The predicted amino acid sequence disclosed herein for s195_10 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol.
The predicted s195_10 protein demonstrated at least some similarity to sequences 3 0 identified as AF009301 (TEB4 protein [Homo sapiens]), X76715 (SSM4 gene product [Saccharomyces cerevisiae]), 246861 (Ssm4p [Saccharomyces cerevisiae]), and (Ssm4p [Saccharomyces cerevisiae]). Based upon sequence similarity, s195_10 proteins and each similar protein or peptide may share at least some activity. The TopPredII

computer program predicts eleven additional potential transmembrane domains within the s195_10 protein sequence, centered around amino acids 130, 170, 210, 260, 320, 470, 520, 560, 600, 650, and 690 of SEQ ID N0:20, respectively. The nucleotide sequence of s195_10 indicates that it may contain a simple GAA repeat region.
Deposit of Clones Clones dol5_4, dx290_1, ek390 4, er471 7, fs40_3, ga63_6, gm335 4, hy370 9, ie47_4, and s195_10 were deposited on June 19, 1997 with the American Type Culture Collection (10801 University Boulevard, Manassas, Virginia 20110-2209 U.S.A.) as an original deposit under the Budapest Treaty and were given the accession number ATCC
98468, 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), and the term of the deposit will comply with 37 C.F.R. ~
1.806.
Each clone has been transfected into separate bacterial cells (E. coli) in this composite deposit. Each clone can be removed from the vector in which it was deposited by performing an EcoRI/NotI digestion (5' site, EcoRI; 3' site, NotI) to produce the appropriate fragment for such clone. Each clone was deposited in either the pED6 or pNOTs vector depicted in Figures 1A and 1B, respectively. The pED6dpc2 vector 2 0 {"pED6") was derived from pED6dpc1 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 CIaI site. In some instances, the deposited clone can become "flipped"
2 5 (i.e., in the reverse orientation) in the deposited isolate. In such instances, the cDNA insert can still be isolated by digestion with EcoRI and NotI. However, NotI will then produce the 5' site and EcoRI will produce the 3' site for placement of the cDNA in proper orientation for expression in a suitable vector. The cDNA may also be expressed from the vectors in which they were deposited.
3 0 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 an oligonucleotide probe that was used to isolate or to sequence each full-length clone is identified below, and should be most reliable in isolating the clone of interest.
Clone Probe Seduence dol5 4 SEQ ID N0:21 dx290 1 SEQ ID N0:22 ek390 4 SEQ ID N0:23 er471 7 SEQ ID N0:24 fs40_3 SEQ ID N0:25 ga63_6 SEQ ID N0:26 gm335 4 SEQ ID N0:27 hy370 9 SEQ ID N0:28 ie47 4 SEQ ID N0:29 s195 10 SEQ ID N0:30 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-2 0 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;
2 5 (b) It should be designed to have a Tm 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-3zP 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 3 0 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 ul of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L-broth containing ampicillin at 100 ug/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 ~g/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 NaCI/liter, 88.2 g Na citrate/liter, adjusted to pH 7.0 with NaOH) containing 0.5% SDS,100 ug/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.
2 0 A third wash with O.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 2 5 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, 3 0 as described in H.U. Saragovi, et al., Bio/Technology 0 773-778 (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 forms) 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 sequences) of the mature forms) 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 2 0 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.
2 5 Organisms that have enhanced, reduced, or modified expression of the genes) 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. 25(7): 250-254; Lavarosky et al., 1997, 3 0 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 genes) 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 B1, incorporated by reference herein).
In addition, organisms are provided in which the genes) corresponding to the polynucleotide sequences disclosed herein have been partially or completely inactivated, through insertion of extraneous sequences into the corresponding genes) 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 products) of the corresponding gene(s).
2 0 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 2 5 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%
3 0 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 homologues 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. Preferably, polynucleotide species homologues have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90%
identity) with the given polynucleotide, and protein species homologues have at least 30%
sequence identity (more preferably, at least 45% identity; most preferably at least 60%
identity) with the given protein, where sequence identity is determined by comparing the nucleotide sequences of the polynucleotides or the amino acid sequences of the proteins when aligned so as to maximize overlap and identity while minimizing sequence gaps.
Species homologues may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from the desired species. Preferably, species homologues are those isolated from mammalian species. Most preferably, species homologues are those isolated from certain mammalian species such as, for example, Pan troglodytes, Gorilla gorilla, Pongo pygmaeus, Hylobates concolor, Macaca mulatta, Papio papio, Papio hamadryas, Cercopithecus aethiops, Cebus capucinus, 2 0 Aotus trivirgatus, Sanguinus Oedipus, Microcebus murinus, Mus musculus, Rattus norvegicus, Cricetulus griseus, Felis catus, Mustela visors, Canis familiaris, Oryctolagus cuniculus, Bos taurus, Ovis cries, Sics scrofa, and E9uus caballus, for which genetic maps have been created allowing the identification of syntenic relationships between the genomic organization of genes in one species and the genomic organization of the related genes in another species 2 5 (O'Brien and Seuanez, 1988, Ann. Rev. Genet. 22: 323-351; O'Brien et al., 1993, Nature Genetics 3:103-112; Johansson et al., 1995, Genomics 25: 682-690; Lyons et al.,1997, Nature Genetics 15: 47-56; O'Brien et al.,1997, Trends in Genetics 13(10): 393-399;
Carver and Stubbs, 1997, Genome Research 7:1123-1137; all of which are incorporated by reference herein).
The invention also encompasses allelic variants of the disclosed polynucleotides 3 0 or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotides which also encode proteins which are identical or have significantly similar sequences to those encoded by the disclosed polynucleotides. Preferably, allelic variants have at least 60% sequence identity (more preferably, at least 75% identity; most preferably at least 90%

WO 98/5797b PCT/US98/12516 identity) with the given polynucleotide, where sequence identity is determined by comparing the nucleotide sequences of the polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps. Allelic variants may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from individuals of the appropriate species.
The invention also includes polynucleotides with sequences complementary to those of the polynucleotides disclosed herein.
The present invention also includes polynucleotides that hybridize 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.

StringencyPolynucleotideHybridHybridization TemperatureWash ConditionHybrid Lengthand Temperature (bp)~ Buffer' and Buffer' A DNA:DNA Z 50 65C; lxSSC -or- 65C; 0.3xSSC
42C; lxSSC, 50% formamide B DNA:DNA <50 TB*; lxSSC TB*; lxSSC

C DNA:RNA z 50 67C; lxSSC -or- 67C; 0.3xSSC
45C; lxSSC, 50% formamide D DNA:RNA <50 Tp*; lxSSC To*; lxSSC

E RNA:RNA z 50 70C; lxSSC -or- 70C; 0.3xSSC
50C; lxSSC, 50% formamide F RNA:RNA <50 TF*; lxSSC TF*; lxSSC

G DNA:DNA s 50 65C; 4xSSC -or- 65C; lxSSC
42C; 4xSSC, 50% formamide H DNA:DNA <50 T,,*; 4xSSC TH*; 4xSSC

I DNA:RNA z 50 67C; 4xSSC -or- 67C; lxSSC
45C; 4xSSC, 50% formamide J DNA:RNA <50 T~*; 4xSSC T~*; 4xSSC

K RNA:RNA s 50 70C; 4xSSC -or- 67C; lxSSC
50C; 4xSSC, 50% formamide L RNA:RNA <50 T~*; 2xSSC T~*; 2xSSC

M DNA:DNA s 50 50C; 4xSSC -or- 50C; 2xSSC
40C; 6xSSC, 50% formamide N DNA:DNA <50 TN*; 6xSSC T"*; 6xSSC

O DNA:RNA s 50 55C; 4xSSC -or- 55C; 2xSSC
42C; 6xSSC, 50% formamide P DNA:RNA <50 T,~*; 6xSSC TP*; 6xSSC

Q RNA:RNA z 50 60C; 4xSSC -or- 60C; 2xSSC
45C; 6xSSC, 50% formamide 2 R RNA:RNA <50 TR*; 4xSSC TR*; 4xSSC

$: The hybrid length is that anticipated for the hybridized regions) of the hybridizing 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 2 5 hybrid length can be determined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity.
t: SSPE (lxSSPE is 0.15M NaCI, lOmM NaHZPO" and 1.25mM EDTA, pH 7.4) can be substituted for SSC
(lxSSC is 0.15M NaCI and lSmM sodium citrate) in the hybridization and wash buffers; washes are performed for 15 nvnutes after hybridization is complete.
3 0 *TB - TR: The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10°C less than the melting temperature (Tm) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, Tm(°C) = 2(# of A + T bases) + 4(# of G +
C bases). For hybrids between 18 and 49 base pairs in length, Tm(°C) =
81.5 + 16.6{log,o[Na']) + 0.41(%G+C) {600/N), where N is the number of bases in the hybrid, and [Na'] is the concentration of sodium ions in the 3 5 hybridization buffer ([Na'] for IxSSC = 0.165 M).

Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E.F. Fritsch, and T. Maniatis, 1989, Moleca~Iar 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. 9 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 2 0 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 2 5 protein. Mammalian host cells include, for example, monkey COS cells, Chinese Hamster Ovary (CHO) cells, human kidney 293 cells, human epidermal A431 cells, human Co1o205 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.
3 0 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 S 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."
I5 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 2 0 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 Sepharose~; one or more steps involving hydrophobic interaction chromatography using such resins as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography.
2 5 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 3 0 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 Rl'-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 2 0 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 2 5 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 3 0 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"
2 0 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 2 5 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, those described in Gyuris et al., 1993, Cell 75: 791-803 and in Rossi et al., 1997, Proc. Natl. Acad. Sci. USA 94: 8405-8410, all of which are incorporated by reference herein) to identify polynucleotides encoding the other protein with which binding occurs or to 3 0 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 2 0 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 2 5 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 3 0 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, RBS, DA1, 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 y, Schreiber, R.D. In Current Protocols in 2 0 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, 2 5 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 3 0 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- oar 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 (SCID)), 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 2 0 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 2 5 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, 3 0 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 2 0 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 2 5 with a monomeric form of a peptide having an activity of another B
lymphocyte antigen (e.g., B7-l, 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 3 0 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. Nati. Acad.
Sci USA, 89:11102-11105 (1992). In addition, marine 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 2 0 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 2 5 human autoimmune diseases. Examples include marine experimental autoimmune encephalitis, systemic lupus erythmatosis in MRL/lpr/lpr mice or NZB hybrid mice, marine autoimmune collagen arthritis, diabetes mellitus in NOD mice and BB
rats, and marine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York,1989, pp. 840-856).
3 0 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.
2 0 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 B7-1-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 2 5 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 antigens) 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
3 0 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 a chain protein and microglobulin protein or an MHC class II a chain protein and an MHC class II
~i 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.
2 0 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.
2 5 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.
3 0 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; Macatorua 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.
2 0 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.
2 5 Hematopoiesis Re ug lating Act 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 3 0 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) WO 98/57976 PCT/US98/1251b 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 2 0 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 2 5 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, 3 0 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 2 0 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 2 5 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 3 0 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, Huntingtori s disease, amyotrophic lateral sclerosis, and Shy-Drager 2 0 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.
2 5 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, 3 0 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 scarnng 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.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for tissue generation activity include, without limitation, those described in: International Patent Publication No. W095/16035 (bone, cartilage, tendon);
International Patent Publication No. W095/05846 (nerve, neuronal);
International Patent Publication No. W091/07491 (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, lnc., 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 2 0 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 a family, may be useful as a contraceptive based on the ability of inhibins to decrease fertility in female mammals 2 5 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-(i 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, 3 0 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 ActivitX
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 2 0 of cells can be readily determined by employing such protein or peptide in any known assay for cell chemotaxis.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Assays for chemotactic activity (which will identify proteins that induce or prevent 2 5 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 3 0 Publishing Associates and Wiley-Interscience (Chapter 6.22, 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 2 0 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 2 5 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.
3 0 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-Inflammatonr 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 2 0 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 Supyressor Activity Cadherins are calcium-dependent adhesion molecules that appear to play major 2 S 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.
3 0 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 2 0 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 2 5 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 3 0 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 poly-nucleotides 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.
2 0 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, 2 5 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, 3 0 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, 2 0 IL-12, IL-13, IL-14, IL-15, IFN, TNFO, TNF1, TNF2, G-CSF, Meg-CSF, thrombopoietin, stem cell factor, and erythropoietin. The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, 2 5 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.
3 0 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 proteins) 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 antigens) 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, saporun, bile acids, 2 0 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 2 5 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 3 0 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 2 0 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 2 5 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.
3 0 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 lxg to about 100 mg (preferably about 0.lng to about 10 mg, more preferably about 0.1 ug 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 2 0 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 2 5 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. T'he peptide immunogens additionally may contain a cysteine residue at the 3 0 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 2 0 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 2 5 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 3 0 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, polyethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and polyvinyl 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.
2 0 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-a and TGF-Vii), and insulin-like growth factor (IGF).
2 5 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 3 0 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.
Treacy, Maurice Spaulding, Vikki Agostino, Michael J.
Hooves, Steven H.
Fechtel, Kim (ii) TITLE OF INVENTION: SECRETED PROTEINS AND POLYNUCLEOTIDES
ENCODING THEM
(iii) NUMBER OF SEQUENCES: 30 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Genetics Institute, Inc.
(B) STREET: 87 CambridgePark Drive (C) CITY: Cambridge 2 S (D) STATE: MA
(E) COUNTRY: U.S.A.
(F) ZIP: 02140 (v) COMPUTER READABLE FORM:
3 0 (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0, Version #1.30 3 S (vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
4O (viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Sprunger, Suzanne A.
(B) REGISTRATION NUMBER: 41,323 (ix) TELECOMMUNICATION INFORMATION:
4 5 (A) TELEPHONE: (617) 498-8284 (B) TELEFAX: (617) 876-5851 (2) INFORMATION FOR SEQ ID N0:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1748 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID
N0:1:

TTTTGATCAC CATACTGAAG AGGATATAGA

O

O

O

O

GTTTTTACGT CATTACAAGC TGAGTAAAATTCCTTCTGAT GATGTTATAA A,~~i~AAAAAAA1740 (2) INFORMATION FOR SEQ
ID N0:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 472 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE
DESCRIPTION:
SEQ
ID
N0:2:

Met GluSerGln PheLeuArg AspThrIle CysGlu GluSerLeuArg Glu LysLeuGln AspGlyArg IleThrIle ArgGlu PhePheIleLeu Leu GlnValHis IleLeuIle GlnLysPro ArgGln SerAsnLeuPro Gly AsnPheThr ValAsnThr ProProThr ProGlu AspLeuMetLeu Ser GlnTyrVal TyrArgPro LysIleGln IleTyr ArgGluAspCys Glu AlaArgArg GlnLysIle GluGluLeu LysLeu SerAlaSerAsn Gln AspLysLeu LeuValAsp IleAsnLys AsnLeu TrpGluLysMet Arg HisCysSer AspLysGlu LeuLysAla PheGly IleTyrLeuAsn Lys IleLysSer CysPheThr LysMetThr LysVal PheThrHisGln Gly LysValAla LeuTyrGly LysLeuVal GlnSer AlaGlnAsnGlu Arg Glu LysLeuGln IleLysIle AspGlu MetAspLys IleLeuLys Lys Ile AspAsnCys LeuThrGlu MetGlu ThrGluThr LysAsnLeu Glu Asp GluGluLys AsnAsnPro ValGlu GluTrpAsp SerGluMet Arg Ala AlaGluLys GluLeuGlu GlnLeu LysThrGlu GluGluGlu Leu Gln ArgAsnLeu LeuGluLeu GluVal ProLysGlu GlnThrLeu Ala Gln IleAspPhe MetGlnLys GlnArg AsnArgThr GluGluLeu Leu Asp GlnLeuSer LeuSerGlu TrpAsp ValValGlu TrpSerAsp Asp Gln AlaValPhe ThrPheVal TyrAsp ThrIleGln LeuThrIle Thr Phe GluGluSer ValValGly PhePro PheLeuAsp LysArgTyr 3 Arg Lys IleValAsp ValAsnPhe GlnSer LeuLeuAsp GluAspGln Ala Pro ProSerSer LeuLeuVal HisLys LeuIlePhe GlnTyrVal Glu Glu LysGluSer TrpLysLys ThrCys ThrThrGln HisGlnLeu Pro Lys MetLeuGlu GluPheSer LeuVal ValHisHis CysArgLeu Leu Gly GluGluIle GluTyrLeu LysArg TrpGlyPro AsnTyrAsn 4 Leu Met AsnIleAsp IleAsnAsn AsnGlu LeuArgLeu LeuPheSer Ser Ser AlaAlaPhe AlaLysPhe GluIle ThrLeuPhe LeuSerAla Tyr Tyr ProSerVal ProLeuPro SerThr IleGlnAsn HisValGly Asn Thr SerGlnAsp AspIleAla ThrIle LeuSerLys ValProLeu Glu Asn Asn Tyr Leu Lys Asn Val Val Lys Gln Ile Tyr Gln Asp Leu Phe Gln Asp Cys His Phe Tyr His (2) INFORMATION FOR SEQ ID N0:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2298 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
2 (xi) SEQUENCE DESCRIPTION:
O SEQ ID N0:3:

O

O

TCACTCTTGC rf~~AAAAAAAA AAAAAAAAAAAAAAAAAAAA P~~4AAAAAAAP,~~ 2 (2) INFORMATION FOR SEQ ID
N0:4:

(i) SEQUENCE CHARACTERISTIC S:

(A) LENGTH: 672 amino acids (B) TYPE: amino acid 5O (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE
DESCRIPTION:
SEQ
ID
N0:4:

Met Ser AspProGluGly GluThrLeu ArgSer ThrPhePro SerTyr Met Ala GluGlyGluArg LeuTyrLeu CysGly GluPheSer LysAla Ala Gln SerPheSerAsn AlaLeuTyr LeuGln AspGlyAsp LysAsn Cys Leu ValAlaArgSer LysCysPhe LeuLys MetGlyAsp LeuGlu Arg Ser LeuLysAspAla GluAlaSer LeuGln SerAspPro AlaPhe Cys Lys GlyIleLeuGln LysAlaGlu ThrLeu TyrThrMet GlyAsp Phe Glu PheAlaLeuVal PheTyrHis ArgXaa TyrLysLeu XaaPro 2 Asp Arg GluPheXaaXaa GlyIleGln LysAla GlnGluAla IleAsn Asn Ser ValGlySerPro SerSerIle LysLeu GluAsnLys GlyAsp Leu Ser PheLeuSerLys GlnAlaGlu AsnIle LysAlaGln GlnLys Pro Gln ProMetLysHis LeuLeuHis ProThr LysGlyGlu ProLys Trp Lys AlaSerLeuLys SerGluLys ThrVal ArgGlnLeu LeuGly Glu Leu TyrValAspLys GluTyrLeu GluLys LeuLeuLeu AspGlu Asp Leu IleLysGlyThr MetLysGly GlyLeu ThrValGlu AspLeu Ile Met ThrGlyIleAsn TyrLeuAsp ThrHis SerAsnPhe TrpArg Gln Gln LysProIleTyr AlaArgGlu ArgAsp ArgLysLeu MetGln Glu Lys TrpLeuArgAsp HisLysArg ArgPro SerGlnThr AlaHis Tyr Ile LeuLysSerLeu GluAspIle AspMet LeuLeuThr SerGly 7$

Ser AlaGluGly SerLeuGln LysAla GluLysVal LeuLysLys Val Leu GluTrpAsn LysGluGlu ValPro AsnLysAsp GluLeuVal Gly Asn LeuTyrSer CysIleGly AsnAla GlnIleGlu LeuGlyGln Met Glu AlaAlaLeu GlnSerHis ArgLys AspLeuGlu IleAlaLys Glu Tyr AspLeuPro AspAlaLys SerArg AlaLeuAsp AsnIleGly Arg Val PheAlaArg ValGlyLys PheGln GlnAlaIle AspThrTrp Glu Glu LysIlePro LeuAlaLys ThrThr LeuGluLys ThrTrpLeu Phe His GluIleGly ArgCysTyr LeuGlu LeuAspGln AlaTrpGln Ala Gln AsnTyrGly GluLysSer GlnGln CysAlaGlu GluGluGly Asp 3 0 Ile GluTrpGln LeuAsnAla SerVal LeuValAla GlnAlaGln Val Lys LeuArgAsp PheGluSer AlaVal AsnAsnPhe GluLysAla Leu Glu ArgAlaLys LeuValHis AsnAsn GluAlaGln GlnAlaIle Ile Ser AlaLeuAsp AspAlaAsn LysGly IleIleArg GluLeuArg Lys Thr AsnTyrVal GluAsnLeu LysGlu LysSerGlu GlyGluAla Ser Leu TyrGluAsp ArgIleIle ThrArg GluLysAsp MetArgArg Val Arg AspGluPro GluLysVal ValLys GlnTrpAsp HisSerGlu Asp Glu LysGluThr AspGluAsp AspGlu AlaPheGly GluAlaLeu Gln Ser ProAlaSer GlyLysGln SerVal GluAlaGly LysAlaArg Ser Asp Leu Gly Ala Val Ala Lys Gly Leu Ser Gly Glu Leu Gly Thr Arg Ser Gly Glu Thr Gly Arg Lys Leu Leu Glu Ala Gly Arg Arg Glu Ser Arg Glu Ile Tyr Arg Arg Pro Ser Gly Glu Leu Glu Gln Arg Leu Ser Gly Glu Phe Ser Arg Gln Glu Pro Glu Glu Leu Lys Lys Leu Ser Glu Val Gly Arg Arg Xaa Pro Glu Glu Leu Gly Lys Thr Gln Phe Gly Glu Ile Gly Glu Thr Lys Lys Thr Gly Asn Glu Met Glu Lys Glu Tyr Glu 2 O (2) INFORMATION FOR SEQ ID N0:5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1010 base pairs {B) TYPE: nucleic acid 2 5 {C) STRANDEDNESS: double (D) TOPOLOGY: linear {ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:5:

(2) INFORMATION FOR SEQ ID
N0:6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 205 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE
DESCRIPTION:
SEQ
ID
N0:6:

Met GlySer GlyGlyLeu GlyAla TrpSerAla GlySerSer ValSer Ala PheHis SerThrAsn AlaVal SerValPhe SerLeuSer LeuPhe Arg AlaCys ThrProVal ProAsp ProAlaPro TrpProIle ProCys Arg GlyAla SerGlyLys LysThr GlyArgPro AlaArgAla ArgLeu Arg ArgGly HisProAla SerPro ProThrAla ArgCysLeu ThrCys Phe AlaThr AspSerCys SerGln ProLeuGly AlaSerVal PheGly Val GlyGlu LeuLeuArg AspCys AlaArgPro ArgProPro ThrLeu Lys CysGln HisHisGln HisGln IlePheArg ArgHisThr AlaLeu Arg ThrArg ArgProGly ArgPhe ValSerSer CysIleLys LeuSer Pro CysPro LeuLeuPro LeuPro ProValPhe LeuLeuIle PhePhe 7g Ser Pro Phe Pro Pro Ser Leu Ser Ala Phe Phe Pro Trp Phe Ser Thr Gly Lys Thr Val Pro Leu Pro Pro Cys Leu His Gly Ser Pro Ala His Val Met Leu Pro Ser Leu Phe Phe Val Cys Val Phe Ile (2) INFORMATION FOR SEQ ID N0:7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2409 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear 2 O (ii) MOLECULE TYPE: cDNA
2 (xi) SEQUENCE DESCRIPTION:
5 SEQ ID N0:7:

ACAAACCTTT
CTGGACGGCA

ACTTTTTAAT AATAAAAAGAAAAGTGAAGAGTAAGAGAAA TTGTAAAAAAP~~~.AAP,AAAA2400 AAAAA.AAAA

(2) INFORMATION
FOR SEQ ID N0:8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 400 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION:
SEQ
ID
N0:8:

Met IleCys AspThr AspProGluLeu GlyGlyAla ValGln LeuMet Gly LeuLeu ArgThr LeuValAspPro GluAsnMet LeuAla ThrAla Xaa LysThr XaaLys ThrGluPheLeu GlyPhePhe TyrLys HisCys Met HisVal LeuXaa AlaProLeuLeu AlaAsnThr ThrGlu AspLys 2 Pro SerLys AspAsp PheGlnThrAla GlnLeuLeu AlaLeu ValLeu Glu LeuLeu ThrPhe CysValGluHis HisThrTyr HisIle LysAsn Tyr IleIle AsnLys AspIleLeuArg ArgValLeu ValLeu MetAla Ser LysHis AlaPhe LeuAlaLeuCys AlaLeuArg PheLys ArgLys Ile IleGly LeuLys AspGluPheTyr AsnArgTyr IleMet LysSer 4 Phe LeuPhe GluPro ValValLysAla PheLeuAsn AsnGly SerArg Tyr AsnLeu MetAsn SerAlaIleIle GluMetPhe GluPhe IleArg Val GluAsp IleLys SerLeuThrAla HisValIle GluAsn TyrTrp Lys AlaLeu GluAsp ValAspTyrVal GlnThrPhe LysGly LeuLys Leu ArgPhe GluGln GlnArgGluArg GlnAspAsn ProLys LeuAsp 5 Ser MetArg SerIle LeuArgAsnHis ArgTyrArg ArgAsp AlaArg gl Thr LeuGlu AspGluGlu GluMetTrp PheAsnThr AspGluAsp Asp Met GluAsp GlyGluAla ValValSer ProSerAsp LysThrLys Asn Asp AspAsp IleMetAsp ProIleSer LysPheMet GluArgLys Lys Leu LysGlu SerGluGlu LysGluVal LeuLeuLys ThrAsnLeu Ser Gly ArgGln SerProSer PheLysLeu SerLeuSer SerGlyThr Lys Thr AsnLeu ThrSerGln SerSerThr ThrAsnLeu ProGlySer Pro Gly SerPro GlySerPro GlySerPro GlySerPro GlySerVal Pro Lys AsnThr SerGlnThr AlaAlaIle ThrThrLys GlyGlyLeu Val Gly LeuVal AspTyrPro AspAspAsp GluAspAsp AspGluAsp Glu 3 Asp LysGlu AspThrLeu ProLeuSer LysLysAla LysPheAsp Ser (2) INFORMATION FOR SEQ ID N0:9:
(ij SEQUENCE CHARACTERISTICS:
{A) LENGTH: 951 base pairs {B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
{xi) SEQUENCE DESCRIPTION:
SEQ ID N0:9:

GATTATTTTT TTTAACTAAA GGAAGATAAAATTCTAAAAAF~~?~AAAAAAA A 951 (2) INFORMATION FOR SEQ
ID NO:10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 87 amino acids (B) TYPE: amino acid 3 O (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION:
SEQ
ID
N0:10:

Met ProTrp AspHisGlyGln GlyArgLeu TrpGly SerGluThr Pro Leu LeuSer ThrProSerGln AsnThrLeu ArgVal SerGlyLeu Trp Arg GluTrp GlyGlyArgLys AsnTrpHis LeuPro ArgGluGly Asp Glu ArgPhe AlaLeuIleLeu ArgGluAla SerGlu LysCysPhe Lys Cys ValCys MetXaaGlnAla ValGlySer GlyGly LeuSerXaa Pro Leu ProPro SerPheProLys (2) INFORMATION FOR SEQ ID N0:11:
5 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1899 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE
DESCRIPTION:
SEQ ID
NO:11:

O

O

O

2 TACATAAAATGTTGATCTTC AAAAAAAAAAp~~AAAAAAA 1899 (2) INFORMATION
FOR SEQ
ID N0:12:

(i) S EQUENCE CHARACTERISTICS:

3 (A) LENGTH: 543 acids 0 amino (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear 3 (ii) MOLECULE
5 TYPE:
protein 4 (xi) SEQUENCE
0 DESCRIPTION:
SEQ
ID
N0:12:

Met ArgLys AlaAlaGly PheLeuPro SerLeu LeuLysVal Leu Pro 4 Leu LeuPro AlaProAla AlaAlaGln AspSer ThrGlnAla Ser 5 Leu Thr ProGly ProLeuSer ProThrGlu TyrGlu ArgPhePhe Ala Ser Leu LeuThr ThrTrpLys AlaGluThr ThrCys ArgLeuArg Ala Pro Thr HisGly ArgAsnPro ThrLeuVal GlnLeu AspGlnTyr Glu Cys Asn His Gly Leu Val Pro Asp Gly Ala Val Cys Ser Asn Leu Pro Tyr Ala Ser TrpPheGlu SerPhe CysGlnPheThr HisTyr ArgCysSer Asn His ValTyrTyr AlaLys ArgValLeuCys SerGln ProValSer Ile Leu SerProAsn ThrLeu LysGluIleGlu AlaSer AlaGluVal Ser Pro ThrThrMet ThrSer ProIleSerPro HisPhe ThrValThr Glu Arg GlnThrPhe GlnPro TrpProGluArg LeuSer AsnAsnVal Glu Glu LeuLeuGln SerSer LeuSerLeuGly GlyGln GluGlnAla Pro Glu HisLysGln GluGln GlyValGluHis ArgGln GluProThr Gln Glu HisLysGln GluGlu GlyGlnLysGln GluGlu GlnGluGlu Glu GIn GluGluGlu GlyLys GlnGluGluGly GlnGly ThrLysGlu Gly Arg GluAlaVal SerGln LeuGlnThrAsp SerGlu ProLysPhe His Ser GluSerLeu SerSer AsnProSerSer PheAla ProArgVal Arg Glu ValGluSer ThrPro MetIleMetGlu AsnIle GlnGluLeu Ile Arg SerAlaGln GluIle AspGluMetAsn GluIle TyrAspGlu Asn Ser TyrTrpArg AsnGln AsnProGlySer LeuLeu GlnLeuPro His Thr GluAlaLeu LeuVal LeuCysTyrSer IleVal GluAsnThr Cys Ile IleThrPro ThrAla LysAlaTrpLys TyrMet GluGluGlu Ile Leu GlyPheGly LysSer ValCysAspSer LeuGly ArgArgHis Met Ser ThrCysAla LeuCys AspPheCysSer LeuLys LeuGluGln Cys HisSer AlaSerLeu GlnArgGln GlnCysAsp ThrSerHis Glu Lys ThrPro ValSerPro LeuLeuAla SerGlnSer LeuSerIle Phe Gly AsnGln Gly5erPro GluSerGly ArgPheTyr GlyLeuAsp Val Leu TyrGly LeuHisMet AspPheTrp CysAlaArg LeuAlaThr Gly 15Lys GlyCys AspValArg ValSerGly TrpLeuGln ThrGluPhe Glu Leu SerPhe AspGlyAsp PheProThr LysIleCys AspThrAsp Gln Tyr IleGln ProAsnTyr CysSerPhe LysSerGln GlnCysLeu Tyr Met ArgAsn AsnArgLys ValSerArg MetArgCys LeuGlnAsn Arg Glu ThrTyr AlaLeuSer LeuAlaLys ValArgThr LeuCysPhe Ser 3 Arg TrpSer GluPheSer ThrLeuThr LeuGlyGln PheGly 0 Gln (2) INFORMATI ON FOR ID
SEQ N0:13:

3 (i) SEQUENCE STICS:

(A)LENGTH:72 2 se ba pairs (B)TYPE: acid nucleic {C)STRANDEDNESS: double (D)TOPOLOGY: linear (ii) MOLECULE cDNA
TYPE:

(xi) SEQUENCE
DESCRIPTION:
SEQ ID N0:13:

AGGCGTCCAG

GATACACTAA P,~~~AAAAAAA F~~,AAAAAAAAF,~~~AAAAAAAP~~.AAAAAAAA P,~~~AAAAAAA7 (2) INFORMATION FOR SEQ ID
N0:14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 169 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

2 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE
DESCRIPTION:
SEQ
ID
N0:14:

Met HisLeu AlaArgLeu ValGlySer CysSer LeuLeuLeu LeuLeu 1 s l0 15 Gly AlaLeu SerGlyTrp AlaAlaSer AspAsp ProIleGlu LysVal Ile GluGly IleAsnArg GlyLeuSer AsnAla GluArgGlu ValGly Lys AlaLeu AspGlyIle AsnSerGly IleThr HisAlaGly ArgGlu Val GluLys ValPheAsn GlyLeuSer AsnMet GlySerHis ThrGly Lys GluLeu AspLysGly ValGlnGly LeuAsn HisGlyMet AspLys Val AlaHis GluIleAsn HisGlyIle GlyGln AlaGlyLys GluAla Glu LysLeu GlyHisGly ValAsnAsn AlaAla GlyGlnGly AsnHis gg Gln Ser Gly Ser Ser Ser His Gln Gly Gly Ala Thr Thr Thr Pro Leu Ala Ser Gly Ala Ser Val Asn Thr Pro Phe Ile Asn Leu Pro Ala Leu Trp Arg Ser Val Ala Asn Ile Met Pro (2) INFORMATION
FOR
SEQ
ID N0:15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1240 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear 2 (ii) MOLECULE TYPE: cDNA
O

2 (xi) SEQUENCE DESCRIPTION:
5 SEQ ID N0:15:

GATCTTGTAC ATGTATTAAA AACTTAAATT AAATGCATTCAAGTTAAAAA F,~~AAAAAAA.A1200 P,F~~AA.A.AAAA F,F~A.AAAAAAA A,F~~AAAAAAA 12 p,~I~~AAAAAAA 4 (2) INFORMATION FOR SEQ ID N0:16:

1 (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 330 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE
DESCRIPTION:
SEQ
ID
N0:16:

Met Ala PheLeuPro SerTrp ValCysVal LeuValGly SerPheSer Ala Ser LeuAlaGly ThrSer AsnLeuSer GluThrGlu ProProLeu Trp Lys GluSerPro GlyGln LeuSerAsp TyrArgVal GluAsnSer Met Tyr IleIleAsn ProTrp ValTyrLeu GluArgMet GlyMetTyr Lys Ile IleLeuAsn GlnThr AlaArgTyr PheAlaLys PheAlaPro Asp Asn GluGlnAsn IleLeu TrpGlyLeu ProLeuGln TyrGlyTrp Gln Tyr ArgThrGly ArgLeu AlaAspPro ThrArgArg ThrAsnCys Gly Tyr GluSerGly AspHis MetCysIle SerValAsp SerTrpTrp Ala Asp LeuAsnTyr PheLeu SerSerLeu ProPheLeu AlaAlaVal 5 Asp Ser GlyVaiMet GlyIle SerSerAsp GlnValArg LeuLeuPro Pro Pro AsnGlu ArgLysPhe CysTyrAsp ValSerSerCys Arg Lys Ser Ser ProGlu ThrMetAsn LysTrpAsn ThrPheTyrGln Tyr Phe Leu Gln ProPhe SerLysPhe AspAspLeu LeuLysTyrLeu Trp Ser Ala Ala ThrSer ThrLeuAla AspAsnIle LysSerPheGlu Asp His Arg Tyr TyrTyr SerLysAla G1uAlaHis PheGluArgSer Trp Asp Val Leu ValAsp HisLeuAla AlaValLeu PheProThrThr Leu Ala Ile Arg TyrLys PheGlnLys GlyMetPro ProArgIleLeu Leu Ser Asn Thr ValAla ProPheIle SerAspPhe ThrAlaPheGln Asn Asp Val Val ValLeu LeuAsnMet LeuAspAsn ValAspLysSer Ile Leu 3 Gly Tyr CysThr GluLysSer AsnValTyr ArgAspHisSer Glu 0 Leu Ser Ser ArgSer TyrGlyAsn AsnSer Ser (2) FORSEQ ID :
INFORMATION N0:17 (i) SEQUENCE TERISTICS:
CHARAC

(A) 61 pairs LENGTH: base (B) nucleicacid TYPE:

(C) double STRANDEDNESS:

(D) linear TOPOLOGY:

(ii) MOLECULE cDNA
TYPE:

(xi) SEQUENCE DESCRIPTION: SEQ ID N0:17:

TAAATAAACA CCTGTGTGTT TAAGAAAAAAP,AAAAAAAAAA 2261 (2) INFORMATION FOR SEQ
ID N0:18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 180 amino acids (B) TYPE. amino acid 2 (C) STRANDEDNESS:
O

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE N0:18:
DESCRIPTION:
SEQ
ID

3 Met AlaSer ThrIleSer AlaTyr LysGluLysMet LysGlu LeuSer Val LeuSer LeuIleCys SerCys PheTyrThrGln ProHis ProAsn Thr ValTyr GlnTyrGly AspMet GluValLysGln LeuAsp LysArg Ala SerGly GlnSerPhe GluVal IleLeuLysSer ProSer AspLeu Ser ProGlu SerProMet LeuSer SerProProLys LysLys AspThr 4 Ser LeuGlu GluLeuGln LysArg LeuGluAlaAla GluGlu ArgArg Lys ThrGln GluAlaGln ValLeu LysGlnLeuAla GluArg ArgGlu His GluArg GluValLeu HisLys AlaLeuGluGlu AsnAsn AsnPhe Ser ArgGln AlaGluGlu LysLeu AsnTyrLysMet GluLeu SerLys Glu Ile Arg Glu Ala His Leu Ala Ala Leu Arg Glu Arg Leu Arg Glu Lys Glu Leu His Ala Ala Glu Val Arg Arg Asn Lys Glu Gln Arg Glu Glu Met Ser Gly (2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3109 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:19:

WO

GGTTTGGGAT

GTTGGGTATA

AAATTTCATA

TTAGTGGTGG

TCCTTGGAAA

GGTACTACCA

TTCATTCTAT

AATGATCCAG

CGAAGATTTA

CCTATACGTA

AGTGATGCTC

GCATTACTCG

GTGACCGCCG

GAAAACAGTG

CCTGTGGTGG

O

GTTGGCTTTC

GTCTTCATGT

GCTGCTTGTG

TGGATGCCTC

ATGAAGACTT

CTGTTTGAGC

TATCCATGGC

TTGATGGGTC

CGGAACATTG

SO

TTGCTTTCCC

ACTGCGGAAA

GTATTGATGG

ATAAAGTAGT

TCTCTTKGGA

CTTGTGTTCT

CTGACATKAC

AACCTTGGAT

TP~AAAP~P.A

(2) INFORMATION FOR SEQ ID N0:20:

(i) SEQUENCE CHARACTERISTICS:

2 (A) LENGTH: 750 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear 2 (ii) MOLECULE TYPE: protein 3 (xi) SEQUENCE DESCRIPTION: N0:20:
O SEQ
ID

Met GlyGlu HisGlnPhe GlyTrp SerMetLeu ProHisArg SerMet 3 Leu ArgGly IleThrLys MetArg LeuGlnGln GluGluMet ValGln Lys MetLeu LeuLeuIle SerLeu LeuThrHis GlnLeuArg ThrGln Trp TrpGly LysThrLeu MetPro ArgMetThr ArgGlnLys ArgArg Arg ArgThr MetArgArg LysMet ThrLeuVal TrpArgMet AlaAla Asp AlaAsn AsnGlyAla GlnAsp AspMetAsn TrpAsnAla LeuGlu g5 90 95 5 Trp AspArg AlaAlaGlu GluLeu ThrTrpGlu ArgMetLeu GlyLeu Asp GlySer LeuValPhe LeuGlu HisValPhe TrpValVal SerLeu Asn Thr Leu Phe Ile Leu Val Phe Ala Phe Cys Pro Tyr His Ile Gly His Phe Ser Leu Val Gly Leu Gly Phe Glu Glu His Val Gln Ala Ser His Phe Glu Gly Leu Ile Thr Thr Ile Val Gly Tyr Ile Leu Leu Ala Ile Thr Leu Ile Ile Cys His Gly Leu Ala Thr Leu Val Lys Phe His Arg Ser Arg Arg Leu Leu Gly Val Cys Tyr Ile Val Val Lys Val Ser Leu Leu Val Val Val Glu Ile Gly Val Phe Pro Leu Ile Cys Gly Trp Trp Leu Asp Ile Cys Ser Leu Glu Met Phe Asp Ala Thr Leu Lys Asp Arg Glu Leu Ser Phe Gln Ser Ala Pro Gly Thr Thr Met Phe Leu His 2 5 Trp Leu Val Gly Met Val Tyr Val Phe Tyr Phe Ala Ser Phe Ile Leu Leu Leu Arg Glu Val Leu Arg Pro Gly Val Leu Trp Phe Leu Arg Asn Leu Asn Asp Pro Asp Phe Asn Pro Val Gln Glu Met Ile His Leu Pro Ile Tyr Arg His Leu Arg Arg Phe Ile Leu Ser Val Ile Val Phe Gly Ser Ile Val Leu Leu Met Leu Trp Leu Pro Ile Arg Ile Ile Lys Ser 4 0 Val Leu Pro Asn Phe Leu Pro Tyr Asn Val Met Leu Tyr Ser Asp Ala Pro Val Ser Glu Leu Ser Leu Glu Leu Leu Leu Leu Gln Val Val Leu Pro Ala Leu Leu Glu Gln Gly His Thr Arg Gln Trp Leu Lys Gly Leu Val Arg Ala Trp Thr Val Thr Ala Gly Tyr Leu Leu Asp Leu His Ser Tyr Leu Leu Gly Asp Gln Glu Glu Asn Glu Asn Ser Ala Asn Gln Gln Val Asn Asn Asn Gln His Ala Arg Asn Asn Asn Ala Ile Pro Val Val Gly Glu GlyLeuHis Ala His AlaIle LeuGlnGln GlyGly Ala Gln Pro Val GlyPheGln ProTyrArg ArgProLeu AsnPhePro LeuArg Ile Phe LeuLeuIle ValPheMet CysIleThr LeuLeuIle AlaSer Leu Ile CysLeuThr LeuProVal PheAlaGly ArgTrpLeu MetSer Phe Trp ThrGlyThr AlaLysIle HisGluLeu TyrThrAla AlaCys Gly Leu TyrValCys TrpLeuThr IleArgAla ValThrVal MetVal Ala Trp MetProGln GlyArgArg ValIlePhe GlnLysVal LysGlu Trp Ser LeuMetIle MetLysThr LeuIleVal AlaValLeu LeuAla Gly Val ValProLeu LeuLeuGly LeuLeuPhe GluLeuVal IleVal 3 Ala Pro LeuArgVal ProLeuAsp GlnThrPro LeuPheTyr ProTrp Gln Asp TrpAlaLeu GlyValLeu HisAlaLys IleIleAla AlaIle Thr Leu MetGlyPro GlnTrpTrp LeuLysThr ValIleGlu GlnVal Tyr Ala AsnGlyIle ArgAsnIle AspLeuHis TyrIleVal ArgLys Leu Ala AlaProVal IleSerVal LeuLeuLeu SerLeuCys ValPro 4 Tyr Val IleAlaSer GlyValVal ProLeuLeu GlyValThr AlaGlu Met Gln Asn Val HisArg IleTyr PheLeu MetVal Leu Arg Pro Leu Val Val Leu Ala Leu Phe Arg Arg Met Ile Ser Gln Gln Val Phe Lys Leu Ile Asn Tyr Lys Asp Glu Lys His Tyr Leu Xaa Gly Gln Xaa Leu Gly Glu Leu Arg Thr Glu Ile Trp Ala Asn Lys Ala His Leu His His Leu His Ser His Pro Lys Asn Lys Val Val Val Ser Thr Thr (2) INFORMATION FOR SEQ ID N0:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:

2 5 (2) INFORMATION FOR SEQ ID N0:22:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid 3 0 (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:22:

(2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"

(xi) SEQUENCE DESCRIPTION: SEQ ID N0:23:

S (2) INFORMATION FOR SEQ ID N0:24:
(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 N0:24:

(2) INFORMATION FOR SEQ ID N0:25:
2 5 (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 N0:25:

(2) INFORMATION FOR SEQ ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs 4 S (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid 5 0 (A) DESCRIPTION: /desc = "oligonucleotide"
5 5 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:26:

(2) INFORMATION FOR SEQ ID N0:27:
(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 N0:27:

(2) INFORMATION FOR SEQ ID N0:28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs 2 5 (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid 3 0 (A) DESCRIPTION: /desc = "oligonucleotide"
3 5 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:28:

(2) INFORMATION FOR SEQ ID N0:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single 4 5 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:29:

lUl (2) INFORMATION FOR SEQ ID N0:30:
(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 N0:30:

Claims

What is claimed is:

1. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 185 to nucleotide 1600;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 1403 to nucleotide 1600;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 1 to nucleotide 850;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone do15_4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone do15_4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone do15_4 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone do15_4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:2;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

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

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

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

5. A process for producing a protein encoded by the polynucleotide of claim 2, which process comprises:
(a) growing a culture of the host cell of claim 3 in a suitable culture medium; and (b) purifying said protein from the culture.

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

7. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:2;
(b) the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 222;
(c) fragments of the amino acid sequence of SEQ ID NO:2 comprising eight consecutive amino acids of SEQ ID NO:2; and (d) the amino acid sequence encoded by the cDNA insert of clone do15_4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

8. The protein of claim 7, wherein said protein comprises the amino acid sequence of SEQ ID NO:2.

9. The protein of claim 7, wherein said protein comprises the amino acid sequence of SEQ ID NO:2 from amino acid 1 to amino acid 222.

10. A composition comprising the protein of claim 7 and a pharmaceutically acceptable carrier.

11. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:1.

12. 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 47 to nucleotide 2065;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:3 from nucleotide 1086 to nucleotide 1848;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone dx290_1 deposited under accession number ATCC 98468;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone dx290_1 deposited under accession number ATCC 98468;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone dx290_1 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:4;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:4 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:4;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (l) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(i).

13. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:4;
(b) the amino acid sequence of SEQ ID NO:4 from amino acid 312 to amino acid 600;

(c) fragments of the amino acid sequence of SEQ ID NO:4 comprising eight consecutive amino acids of SEQ ID NO:4; and (d) the amino acid sequence encoded by the cDNA insert of clone dx290_1 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

15. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 107 to nucleotide 724;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 218 to nucleotide 724;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 536 to nucleotide 866;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ek390_4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ek390_4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ek390_4 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ek390_4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:6;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

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

16. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:6;
(b) the amino acid sequence of SEQ ID NO:6 from amino acid 6 to amino acid 92;
(c) fragments of the amino acid sequence of SEQ ID NO:6 comprising eight consecutive amino acids of SEQ ID NO:6; and (d) the amino acid sequence encoded by the cDNA insert of clone ek390_4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

18. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 31 to nucleotide 1230;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 289 to nucleotide 1230;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 344 to nucleotide 1119;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone er471_7 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone er471_7 deposited under accession number ATCC 98468;

(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone er471_7 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone er471_7 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:8;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

19. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:8;
(b) the amino acid sequence of SEQ ID NO:8 from amino acid 111 to amino acid 363;
(c) fragments of the amino acid sequence of SEQ ID NO:8 comprising eight consecutive amino acids of SEQ ID NO:8; and (d) the amino acid sequence encoded by the cDNA insert of clone er471_7 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

21. 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 62 to nucleotide 322;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 571 to nucleotide 878;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone fs40_3 deposited under accession number ATCC 98468;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone fs40_3 deposited under accession number ATCC 98468;
(f) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone fs40_3 deposited under accession number ATCC
98468;
(g) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone fs40_3 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:10;
(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 (l) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)(i).

22. A protein comprising an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:10;
(b) fragments of the amino acid sequence of SEQ ID NO:10 comprising eight consecutive amino acids of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone fs40_3 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

24. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 43 to nucleotide 1671;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 112 to nucleotide 1671;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 224 to nucleotide 679;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ga63_6 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ga63_6 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ga63_6 deposited under accession number ATCC
98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ga63_6 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:12 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:12;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

25. A protein comprising 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 62 to amino acid 212;
(c) fragments of the amino acid sequence of SEQ ID NO:12 comprising eight consecutive amino acids of SEQ ID NO:12; and (d) the amino acid sequence encoded by the cDNA insert of clone ga63_6 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

26. An isolated gene corresponding to the cDNA sequence of SEQ ID NO:11.

27. 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 17 to nucleotide 523;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 77 to nucleotide 523;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 1 to nucleotide 392;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone gm335_4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone gm335_4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone gm335_4 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone gm335_4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14;

(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:14 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:14;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

28. A protein comprising 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 125;
(c) fragments of the amino acid sequence of SEQ ID NO:14 comprising eight consecutive amino acids of SEQ ID NO:14; and (d) the amino acid sequence encoded by the cDNA insert of clone gm335_4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

30. 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 2 to nucleotide 991;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 62 to nucleotide 991;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 2 to nucleotide 504;

(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone hy370_9 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone hy370_9 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone hy370_9 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone hy370_9 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:16;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

31. A protein comprising 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 167;
(c) fragments of the amino acid sequence of SEQ ID NO:16 comprising eight consecutive amino acids of SEQ ID NO:16; and (d) the amino acid sequence encoded by the cDNA insert of clone hy370_9 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

33. 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 77 to nucleotide 616;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 164 to nucleotide 616;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 1 to nucleotide 415;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone ie47_4 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone ie47_4 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone ie47_4 deposited under accession number ATCC
98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone ie47_4 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:18 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:18;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

34. A protein comprising 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 113;
(c) fragments of the amino acid sequence of SEQ ID NO:18 comprising eight consecutive amino acids of SEQ ID NO:18; and (d) the amino acid sequence encoded by the cDNA insert of clone ie47_4 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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

36. 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 564 to nucleotide 2813;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 705 to nucleotide 2813;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 793 to nucleotide 1628;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone s195_10 deposited under accession number ATCC 98468;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone s195_10 deposited under accession number ATCC 98468;
(g) a polynucleotide comprising the nucleotide sequence of a mature protein coding sequence of clone s195_10 deposited under accession number ATCC 98468;
(h) a polynucleotide encoding a mature protein encoded by the cDNA
insert of clone s195_10 deposited under accession number ATCC 98468;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:20;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:20 having biological activity, the fragment comprising eight consecutive amino acids of SEQ ID NO:20;

(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a)-(j).

37. A protein comprising 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 78 to amino acid 355;
(c) fragments of the amino acid sequence of SEQ ID NO:20 comprising eight consecutive amino acids of SEQ ID NO:20; and (d) the amino acid sequence encoded by the cDNA insert of clone s195_10 deposited under accession number ATCC 98468;
the protein being substantially free from other mammalian proteins.

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