CA2276092A1 - Secreted proteins and polynucleotides encoding them - Google Patents

Abstract

Novel polynucleotides and the proteins encoded thereby are disclosed.

Description

wo 9sr~o69s rcz.~rs9sroosa3 SECRETED PROTEINS AND POLYNUCLEOTIDES ENCODING THEM
This application is a continuation-in-part of Ser. No. 06/:~XX,XXX (converted to a provisional application from non-provisional application Ser. No.
08/780,814), filed January 9, 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) wo 98/30695 PCT/US98/00543 SUMMARY OF THE INVENTION
In one embodiment, the present invention provides a composition comprising an isolated polynucleoHde 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
N0:1 from nucleotide 79 to nucleotide b12;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;
(d) a polynucleotfde encoding the full-length protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2;

2 0 (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:2 having biological activity;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a species homologue of the protein 2 5 of {g) or (h) above ; and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
NO:1 from nucleotide 79 to nucleotide 612; the nucleotide sequence of the full-length 3 0 protein coding sequence of clone BH272 3 deposited under accession number ATCC
98289; or the nucleotide sequence of the mature protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone BH272_3 deposited under accession number ATCC 98289. In yet other preferred wo s rcz.,os~
embodiments, the present invention provides a polynudeotide encoding a protein comprising the amino acid sequence of SEQ ID N0:2 from amino and 13 to amino acid 164.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID NO:1.
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 sequenre of SEQ ID N0:2;
(b) the amino acid sequence of SEQ ID N0:2 from amino acid 13 to amino acid 164;
(c) fragments of the amino acid sequence of SEQ ID N0:2; and (d) the amino acid sequence encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:2 or the amino acid sequence of SEQ ID N0:2 from amino acid 13 to amino acid 164.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
2 0 (a) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:3;
(b) a polynudeotide comprising the nucleotide sequence of SEQ ID
N0:3 from nucleotide 66 to nucleotide 1256;
(c) a polynudeotide comprising the nucleotide sequence of SEQ ID
2 5 N0:3 from nucleotide 1 to nucleotide 332;
(d) a polynudeotide comprising the nucleotide sequence of the full-length protein coding sequence of done BP202_3 deposited under accession number ATCC 98289;
(e) a polynudeotide encoding the full-length protein encoded by the 3 0 cDNA insert of clone BP202_3 deposited under accession number ATCC 98289;
(f) a polynudeotide comprising the nucleotide sequence of the mature protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289;

(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BP202 3 deposited under accession number ATCC 98289;
(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 amino acid sequence of SEQ ID N0:4 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:3 from nucleotide 66 to nucleotide 1256; the nucleotide sequence of SEQ ID
N0:3 from nucleotide 1 to nucleotide 332; the nucleotide sequence of the full-length protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone BP202_3 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of 2 0 clone BP202_3 deposited under accession number ATCC 98289. 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 1 to amino acid 89.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:3.
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 N0:4;
(b) the amino acid sequence of SEQ ID N0:4 from amino acid 1 to 3 0 amino acid 89;
(c) fragments of the amino acid sequence of SEQ ID N0:4; and (d) the amino acid sequence encoded by the cDNA insert of clone BP202_3 deposited under accession number ATCC 98289;

wo ~s~os9s rcr~rs3 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 of SEQ ID N0:4 from amino acid 1 to amino acid 89.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleo#ide comprising the nucleotide sequence of SEQ ID
N0:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 100 to nucleotide 675;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:5 from nucleotide 208 to nucleotide 489;
(d) a polynudeotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CH27 1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequenre of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 9$289;
2 0 (g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CH27 1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:6;
(i) a polynucleotide encoding a protein comprising a fragment of the 2 5 amino acid sequence of SEQ ID N0:6 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (I} a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:5 from nucleotide 100 to nucleotide 675; the nucleotide sequence of SEQ ID
N0:5 from nucleotide 208 to nucleotide 489; the nucleotide sequence of the full-length protein WO 98/30695 PCTlUS98100543 coding sequence of clone CH27 1 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone CI-i27_1 deposited under accession number ATCC 98289. 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 39 to amino acid 130.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
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;
(b) the amino acid sequenre of SEQ ID N0:6 from amino acid 39 to amino acid 130;
(c) fragments of the amino acid sequence of SEQ ID N0:6; and {d) the amino acid sequence encoded by the cDNA insert of clone CI-i27_1 deposited under accession number ATCC 98289;
2 0 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 39 to amino acid 130.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
2 5 (a) a polynucleoHde comprising the nucleotide sequence of SEQ ID
N0:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:7 from nucleotide 168 to nucleotide 1448;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
3 0 N0:7 from nucleotide 440 to nucleotide 942;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289;

WO 98/30695 PCT/US98ro0543 (e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
{f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CI542 2 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:8;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:8 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:7 from nucleotide 168 to nucleotide 1448; the nucleotide sequence of SEQ ID
N0:7 2 0 from nucleotide 440 to nucleotide 941; the nucleotide sequence of the full-length protein coding sequence of clone CI542 2 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of 2 5 clone CI542_2 deposited under accession number ATCC 98289. 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 170 to amino acid 258.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
3 0 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 consisting of:
(a) the amino acid sequence of SEQ ID N0:8;

WO 98!30695 PCTlUS98/00543 (b) the amino acid sequence of SEQ ID N0:8 from amino acid 170 to amino acid 258;
(c) fragments of the amino acid sequence of SEQ ID N0:8; and (d) the amino acid sequence encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID N0:8 or the amino acid sequence of SEQ ID N0:8 from amino acid 170 to amino acid 258.
In one embodiment, the present invention provides a composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleoHde comprising the nucleotide sequence of SEQ ID
N0:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 112 to nucleotide 1212;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 166 to nucleotide 1212;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:9 from nucleotide 704 to nucleotide 1033;
(e) a polynucleotide comprising the nucleotide sequence of the full-2 0 length protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CN483 2 deposited under accession number ATCC 98289;
(g) a polynucleotide comprising the nucleotide sequence of the mature 2 5 protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;
(i) a polynucleotide encoding a protein comprising the amino acid 3 0 sequence of SEQ ID NO:10;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:10 having biological activity;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;

wo 9sr3os9s rcTr~s~oos43 (1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:9 from nucleotide 112 to nucleotide 1212; the nucleotide sequence of SEQ ID
N0:9 from nucleotide 166 to nucleotide 1212; the nucleotide sequence of SEQ ID N0:9 from nucleotide 704 to nucleotide 1033; the nucleotide sequence of the full-length protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone CN483_2 deposited under accession number ATCC 98289.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:9.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:10;
2 0 (b) fragments of the amino acid sequence of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone CN483 2 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins. Preferably such protein comprises the amino acid sequence of SEQ ID NO:10.
2 5 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
3 0 NO:11 from nucleotide 234 to nucleotide 1466;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 1 to nucleotide 367;

WO 98130695 PGT/US98~0543 (d) a polynudeotide comprising the nucleotide sequence of the full-length protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone C01224_2 deposited under accession number ATCC 98289;
(f) a polynudeotide comprising the nucleotide sequence of the mature protein coding sequence of done C01224 2 deposited under accession number ATCC 98289;
(g) a polynudeotide encoding the mature protein encoded by the cDNA insert of done C01224_2 deposited under accession number ATCC 98289;
(h) a polynudeotide encoding a protein comprising the amino acid sequence of SEQ ID N0:12;
(i) a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:12 having biological activity;
(j) a polynudeotide which is an allelic variant of a polynudeotide of (a)-(g) above;
(k) a polynudeotide whidl encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynudeotide capable of hybridizing under stringent conditions 2 0 to any one of the polynudeotides specified in (a)-(i).
Preferably, such polynudeotide comprises the nucleotide sequence of SEQ ID
N0:11 from nucleotide 234 to nucleotide 1466; the nucleotide sequence of SEQ
ID N0:11 from nucleotide 1 to nucleotide 367; the nucleotide sequence of the full-length protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289;
or 2 5 the nucleotide sequence of the mature protein coding sequence of clone C01224_2 deposited under accession number ATCC 98289. In other preferred embodiments, the polynudeotide encodes the full-length or mature protein encoded by the cDNA
insert of clone C01224_2 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynudeotide encoding a protein 3 0 comprising the amino acid sequence of SEQ ID N0:12 from amino acid 1 to amino acid 52.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:11.

wo 9sr~or~s rc~rms3 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;
(b) the amino acid sequence of SEQ ID N0:12 from amino acid 1 to amino acid 52;
(c) fragments of the amino acid sequence of SEQ ID N0:12; and (d) the amino acid sequence encoded by the cDNA insert of clone C01224_2 deposited under accession number ATCC 98289;
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 1 to amino acid 52.
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 845 to nucleotide 1102;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
2 0 N0:13 from nucleotide 828 to nucleotide 1111;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the 2 5 cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequenre of the mature protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the 3 0 cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:14;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:14 having biological activity;

wo 9s~9s pcTivs9a~oosa3 (j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:13 from nucleotide 845 to nucleotide 1102; the nucleotide sequence of SEQ
ID N0:13 from nucleotide 828 to nucleotide 1111; the nucleotide sequence of the full-length protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone CW768_1 deposited under accession number ATCC 98289.
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 consisting of:
2 0 (a) the amino acid sequence of SEQ ID N0:14;
(b) fragments of the amino acid sequence of SEQ ID N0:14; and (c) the amino acid sequence encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins. Preferably such 2 5 protein comprises the amino acid sequence 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;
3 0 (b) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 116 to nucleotide 1081;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:15 from nucleotide 509 to nucleotide 860;

wo 9sr3o69s rc~rrtrs9sioos43 (d) a polynudeotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;
(e) a polynudeotide encoding the full-length protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
(f) a polynudeotide comprising the nucleotide sequence of the mature protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;
(g) a polynudeotide encoding the mature protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:16;
(i) a polynudeotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:16 having biological activity;
(j) a polynudeotide which is an allelic variant of a polynudeotide of (a)-(g) above;
(k) a polynudeotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynudeotide capable of hybridizing under stringent conditions 2 0 to any one of the polynudeotides specified in (a)-(i).
Preferably, such polynudeotide comprises the nucleotide sequence of SEQ ID
N0:15 from nucleotide 116 to nucleotide 1081; the nucleotide sequence of SEQ
ID N0:15 from nucleotide 509 to nucleotide 860; the nucleotide sequence of the full-length protein coding sequence of done CZ268_1 deposited under accession number ATCC 98289;
or the 2 5 nucleotide sequence of the mature protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynudeotide encodes the full-length or mature protein encoded by the cDNA
insert of clone CZ268_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein 3 0 comprising the amino acid sequence of SEQ ID N0:16 from amino acid 153 to amino acid 248.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:15.

wo 9sr3o69s ~~s 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 153 to amino acid 248;
(c) fragments of the amino acid sequence of SEQ ID N0:16; and (d) the amino acid sequence encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
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 153 to amino acid 248.
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 348 to nucleotide 2663;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
2 0 N0:17 from nucleotide 397 to nucleotide 722;
{d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the 2 5 cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the 3 0 cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:18;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID N0:18 having biological activity;

(j) a polynucleotide which is an allelic variant of a polynucieotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (1) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
N0:17 from nucleotide 348 to nucleotide 2663; the nucleotide sequence of SEQ
ID N0:17 from nucleotide 397 to nucleotide 722; the nucleotide sequence of the full-length protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;
or the nucleotide sequence of the mature protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone DH1308_1 deposited under acression number ATCC 98289. 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 19 to amino acid 125.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:17.
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 N0:18;
(b) the amino acid sequence of SEQ ID N0:18 from amino acid 19 to 2 5 amino acid 125;
(c) fragments of the amino acid sequence of SEQ ID N0:18; and (d) the amino acid sequence encoded by the cDNA insert of clone DI-i1308_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins. Preferably such 3 0 protein comprises the amino acid sequence of SEQ ID N0:18 or the amino acid sequence of SEQ ID N0:18 from amino acid 19 to amino acid 125.
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 158 to nucleotide 3268;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 215 to nucleotide 3268;
(d) a polynucleotfde comprising the nucleotide sequence of SEQ ID
N0:19 from nucleotide 55 to nucleotide 379;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;
{f) a polynucleoHde encoding the full-length protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;
(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;
(i) a polynucleoHde encoding a protein comprising the amino acid 2 0 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;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
2 5 (1) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above ; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).
Preferably, such polynucleotide comprises the nucleotide sequence of SEQ ID
3 0 N0:19 from nucleotide 158 to nucleotide 3268; the nucleotide sequence of SEQ ID N0:19 from nucleotide 215 to nucleotide 3268; the nucleotide sequence of SEQ ID
N0:19 from nucleotide 55 to nucleotide 379; the nucleotide sequence of the full-length protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289; or the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289. In other preferred embodiments, the polynucleotide encodes the full-length or mature protein encoded by the cDNA
insert of clone DL185_1 deposited under accession number ATCC 98289. In yet other preferred embodiments, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID N0:20 from amino acid 1 to amino acid 74.
Other embodiments provide the gene corresponding to the cDNA sequence of SEQ
ID N0:19.
In other embodiments, the present invention provides a composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID N0:20;
(b) the amino acid sequence of SEQ ID N0:20 from amino acid 1 to amino acid 74;
(c) fragments of the amino acid sequence of SEQ ID N0:20; and (d) the amino acid sequence encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;
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 2 0 of SEQ ID N0:20 from amino acid 1 to amino acid 74.
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 5 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 3 0 (b) purifying the protein from the culture.
The protein produced according to such methods is also provided by the present invention. Preferred embodiments include those in which the protein produced by such process is a mature form of the protein.

Protein compositions of the present invention may further comprise a pharmaceutically acceptable carrier. Compositions comprising an antibody which specifically reacts with such protein are also provided by the present invention.
Methods are also provided for preventing, treating or ameliorating a medical condition which comprises administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the present invention and a pharmaceutically acceptable earner.
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) can then be determined kom such nucleotide sequence. The amino acid 2 0 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 kame best identifiable with sequence information available at the time of filing.
2 5 As used herein a "secreted" protein is one which, when expressed in a suitable host cell, is transported across or through a membrane, including transport as a result of signal sequences in its amino acid sequence. "Secreted" proteins include without limitation proteins secreted wholly (e.g., soluble proteins) or partially (e.g. , receptors) kom the cell in which they are expressed. "Secreted" proteins also include without limitation proteins 3 0 which are transported across the membrane of the endoplasmic reticulum.
Clone "BH272 3"
A polynucleotide of the present invention has been identified as clone 'BH272 3".
BH272_3 was isolated kom a human adult ovary (PA-1 teratocarcinoma, pooled retinoic-acid-treated, activin-treated, and untreated 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. BH272_3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BH272 3 protein").
The nucleotide sequence of BH272_3 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 BH272_3 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:2.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone BH272_3 should be approximately 1200 bp.
The nucleotide sequence disclosed herein for BH272 3 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BH272_3 demonstrated at least some similarity with sequences identified as AA030161 (mh86cll.rl Soares mouse placenta 4NbMP13.5 14.5 Mus ' musculus), AA088849 (z190d02.s 1 Stratagene colon (#937204) Homo Sapiens cDNA clone 511875 3'), AA099884 (z190d02.r 1 Stratagene colon (#937204) Homo Sapiens cDNA
clone 511875 5' similar to TR 6609419 6609419 CHROMOSOME XII COSMID 8039), 2 0 AA 102282 (zl81 g04.r1 Stratagene colon (#937204) Homo sapiens cDNA clone 5' similar to TR 6609419 6609419 CHROMOSOME XII COSMID 8039), AA 150255 (z107e04.r1 Soares pregnant uterus NbHPU Homo sapiens cDNA clone 491646 5' similar to TR 6609419 6609419 CHROMOSOME XII COSMID 8039), AA150366 (z107b05.r1 Soares pregnant uterus NbHPU Homo sapiens cDNA clone 491601 5' similar to TR
2 5 6609419 6609419 CHROMOSOME XII COSMID 8039), AF004561 {Homo sapiens p21-Arc mRNA, complete CDs), N98300 (yy68c05.r1 Homo sapiens cDNA clone 2?8696 5' similar to PIR:S51388 S51388 hypothetical protein L8039.15 - yeast), 894679 (yq42g I 2.r 1 Homo sapiens cDNA clone 198502 5'), S51388 (hypothetical protein L8039.15 - yeast), T34031 (EST61558 Homo sapiens cDNA 5' end similar to None), 3 0 T56617 (yb35dOl .s 1 Homo Sapiens cDNA clone 73153 3'), W32065 (zb95f09.r1 Soares parathyroid tumor NbHPA Homo Sapiens cDNA clone 320585 5' similar to PIR:S51388 S51388 hypothetical protein L8039.15 - yeast), and W58592 (zd 19d12.r1 Soares fetal heart wo ~os9s rcrrtrs9sroosa3 NbHHI9W Homo Sapiens cDNA clone 341111 5' similar to P1R:S51388 S51388 hypothetical protein L8039.15 - yeast). The predicted amino acid sequence disclosed herein for BH272_3 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BH272_3 protein demonstrated at least some similarity to sequences identified as AF004561 (p21-Arc [Homo Sapiens]) and U19103 (L8039.15 gene product [Saccharomyces cerevisiae]).
Based upon sequence similarity, BH272_3 proteins and each similar protein or peptide may share at least some activity.
Clone "BP202 3"
A polynucleotide of the present invention has been identified as clone "BP202 3".
BP202 3 was isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. BP202_3 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BP202 3 protein') The nucleotide sequence of BP202_3 as presently determined is reported in SEQ
ID N0:3. What applicants presently believe to be the proper reading frame and the 2 0 predicted amino acid sequence of the BP202 3 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 BP202_3 should be approximately 1800 bp.
The nucleotide sequence disclosed herein for BP202_3 was searched against the 2 5 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. BP202_3 demonstrated at least some similarity with sequences identified as AA112336 {zn68a12.r1 Stratagene HeLa cell s3 937216 Homo sapiens cDNA
clone 563326 5'), AA505145 (aa65f02.s1 NCI_CGAP GCB1 Homo sapiens cDNA clone IMAGE:825819 3'}, and T05406 (Human cell cycle gene MIND. The predicted amino acid 3 0 sequence disclosed herein for BP202_3 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted BP202_3 protein demonstrated at least some similarity to sequences identified as 877417 (Human cell cycle protein minl ). MIN1 is a human-derived cell cycle gene encoding an wo 9sr~os9s anti-mitotic factor, and is involved in the control of the G2 /M phase of the cell cycle.
Based upon sequence similarity, BP202_3 proteins and each similar protein or peptide may share at least some activity.
Clone "CH27 1"
A polynucleotide of the present invention has been identified as clone "CH27 1 ".
CH27_1 was isolated from a human fetal kidney cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CH27 1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CH27_2 protein').
The nucleotide sequence of CH27_1 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 CH27_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:6.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CH27_1 should be approximately 1010 bp.
The nucleotide sequence disclosed herein for CH27_1 was searched against the 2 0 GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CH27_1 demonstrated at least some similarity with sequences identified as AA086876 (mn93c10.r1 Stratagene mouse Tcell 937311 Mus musculus cDNA
clone 551634 5' similar to SW SPC3_CANFA P13679 MICRO50MAL SIGNAL PEPTIDASE
21 KD SUBUNTT), C21419 (HUMGS0009728, Human Gene Signature, 3'-directed cDNA
2 5 sequence), J05069 (Canine 21 kDa signal peptidase subunit mRNA, complete cds), and W54889 (ma31d03.r1 Life Tech mouse brain Mus musculus cDNA clone 312293 5' similar to SW SPC3_CANFA P13679 MICROSOMAL SIGNAL PEPTIDASE 21 KD SUBUNIT).
The predicted amino acid sequence disclosed herein for CH27 1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol.
3 0 The predicted CH27_1 protein demonstrated at least some similarity to sequences identified as J05069 (signal peptidase 21 kDa subunit [Cams familiaris]).
Based upon sequence similarity, CH27 1 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts a potential transmembrane domain within the CH27 1 protein sequence centered around amino acid 47 of SEQ
ID
N0:6.
Clone "CI542 2"
A polynucleotide of the present invention has been identified as clone "CI542 2".
CI542_2 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CI542_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CI542 2 proteiri').
The nucleotide sequence of CI542_2 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 CI542_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:8.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CI542_2 should be approximately 1700 bp.
The nucleotide sequence disclosed herein for CI542_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and 2 0 FASTA search protocols. CI542_2 demonstrated at least some similarity with sequences identified as AA114071 (zn76b06.s1 Stratagene NT2 neuronal precursor 937230 Homo sapiens cDNA clone 564083 3'), AA258839 (zs33h10.r1 NCI CGAP_GCB1 Homo sapiens cDNA clone 1MAGE:687043 5'), 806973 (yf12a09.r1 Homo Sapiens cDNA clone 126616 5'), W20825 (mb9lfOb.r1 Scares mouse p3NMF19.5 Mus musculus cDNA clone 336803 5'), 2 5 W93955 (zd98f03.r1 Scares fetal heart NbHHI9W Homo sapiens cDNA clone 357533 5'), and W94010 {zd98f03.s1 Scares fetal heart NbHHI9W Homo sapiens cDNA clone 3'). Based upon sequence similarity, CI542 2 proteins and each similar protein or peptide may share at least some activity.
3 0 Clone "CN483 2"
A polynucleotide of the present invention has been identified as clone "CN483 2".
CN483 2 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer WO 9&'30695 PCTlUS98/00543 analysis of the amino acid sequence of the encoded protein. CN483_2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "CN483 2 protein").
The nucleotide sequence of CN483_2 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 CN483_2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID NO:10. Amino acids 6 to 18 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 19, or are a transmembrane domain.
The EcoRI/Noti restriction fragment obtainable from the deposit containing clone CN483 2 should be approximately 2000 bp.
The nucleotide sequence disclosed herein for CN483_2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. CN483 2 demonstrated at least some similarity with sequences identified as AA112426 (zm27d08.r1 Stratagene pancreas (#937208) Homo sapiens cDNA
clone 526863 5'), AA475778 (vh23fOl.r1 Soares mouse mammary gland NbMMG Mus musculus), H51262 (yp83b07.s1 Homo Sapiens cDNA clone 194005 3'), N57441 (yw88b09.r1 Homo sapiens cDNA clone 259289 5'), W02460 (za47cOl.r1 Soares fetal liver spleen 1NFLS Homo Sapiens cDNA clone 295680 5'), and X92871 (X.laevis mIZIVA
for an 2 0 unknown transmembrane protein). The predicted amino acid sequence disclosed herein for CN483_2 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CN483_2 protein demonstrated at least some similarity to sequences identified as X92871 (unknown transmembrane protein [Xenopus laevis]). Based upon sequence similarity, CN483 2 5 proteins and each similar protein or peptide may share at least some activity. The TopPredII computer program predicts four potential transmembrane domains within the CN483 2 protein sequence centered around amino acids 190, 220, 300, and 350 of SEQ ID
N0:10, respectively.
3 0 Clone "C01224 2"
A polynucleotide of the present invention has been identified as clone "C01224 2".
C01224_2 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. C01224 2 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "C01224 2 protein").
The nucleotide sequence of C01224_2 as presently determined is reported in SEQ
ID NO:11. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the C01224 2 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:12.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone C01224_2 should be approximately 2300 bp.
The nucleotide sequence disclosed herein for C01224 2 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. No hits were found in the database.
Clone "CW768 1"
A polynucleotide of the present invention has been identified as clone "CW768 1".
CW768_1 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CW768_1 is a full-length 2 0 clone, including the entire coding sequence of a secreted protein (also referred to herein as "CW768_1 protein").
The nucleotide sequence of CW768_1 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 CW768_1 protein corresponding to the foregoing 2 5 nucleotide sequence is reported in SEQ ID N0:14.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone CW768_1 should be approximately 1200 bp.
The nucleotide sequence disclosed herein for CW768_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and 3 0 FASTA search protocols. CW768_1 demonstrated at least some similarity with sequences identified as AA425283 (zw46d02.s2 Soares total fetus Nb2HF8 9w Homo Sapiens cDNA
clone 773091 3' similar to contains Alu repetitive element;contains element repetitive element), N27362 (yw52g05.s1 Homo sapiens cDNA clone 255896 3' similar to contains Alu repetitive elementcontains element MERl repetitive element), WO 98/30695 PCT/US98~0543 (Human brain Expressed Sequence Tag EST01598), U52112 (Human Xq28 genomic DNA
in the region of the L1CAM locus containing the genes for neural cell adhesion molecule Ll (L1CAM), arginine-vasopressin receptor (AVPR2}, Cl p115 (Cl)), and 282201 (Human DNA sequence from PAC 345P10 on chromosome 22q12-qter contains ESTs and STS
and polymorphic CA repeat D22S927). The predicted amino acid sequence disclosed herein for CW768_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CW768_1 protein demonstrated at least some similarity to sequences identified as L24521 (transformation-related protein [Homo sapiens]). Based upon sequence similarity, CW768_1 proteins and each similar protein or peptide may share at least some activity.
The nucleotide sequence of CW768_1 indicates that it may contain an Alu repetitive element.
Clone "CZ268 1"
A polynudeotide of the present invention has been identified as clone "CZ268_1".
CZ268_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 identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. CZ268_1 is a full-length 2 0 clone, including the entire coding sequence of a secreted protein (also referred to herein as "CZ268_1 protein").
The nucleotide sequence of CZ268_1 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 CZ268_1 protein corresponding to the foregoing 2 5 nucleotide sequence is reported in SEQ ID N0:16.
The EcoRI/NotI restriction fragment obtainable from the deposit containing done CZ268_1 should be approximately 1500 bp.
The nucleotide sequence disclosed herein for CZ268_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and 3 0 FASTA search protocols. CZ268_1 demonstrated at least some similarity with sequences identified as AA108889 (m156b11.r1 Stratagene mouse testis (#937308) Mus musculus cDNA clone 515997 5' similar to SW:MDP1_PIG P22412 MICROSOMAL DIPEPTIDASE
PRECURSOR), D13138 (Human mRNA for dipeptidase), H10870 (ym06a06.r1 Homo sapiens cDNA clone 47018 5' similar to SP MDPl PIG P22412 MICROSOMAL

wo ~s~os9s rcT~rs9sroos43 DIPEPTIDASE PRECURSOR), J05257 (Homo sapiens (clones MDP4, MDP7) microsomal dipeptidase (MDP) mRNA, complete cds), Q35201 (pDHP2 coding sequence), and (yf59a05.s1 Homo Sapiens cDNA clone 26189 3' similar to SP MDPl RABIT P31429 MICROSOMAL DIPEPTIDASE PRECURSOR). The predicted amino acid sequence disclosed herein for CZ268_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted CZ268_1 protein demonstrated at least some similarity to sequences identified as D13137 (dipeptidase precursor [Homo sapiens]), J05257 (microsomal dipeptidase [Homo sapiens]), and (Microsomal dipeptidase). Microsomal dipeptidase proteins may be associated with brush-border membranes. Based upon sequence similarity, CZ268_1 proteins and each similar protein or peptide may share at least some activity.
The TopPredII computer program predicts a potential transmembrane domain within the CZ268_1 protein sequence centered around amino acid 312 of SEQ ID
N0:16.
Amino acids 151 to 163 of SED ID N0:16 are a putative leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 164, or are a transmembrane domain.
Clone "DH1308 1"
A polynucleotide of the present invention has been identified as clone "DH1308 1".
2 0 DH1308_1 was isolated from a human fetal brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DH1308 1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DH1308_1 protein').
The nucleotide sequence of DH1308_1 as presently determined is reported in SEQ
ID N0:17. What applicants presently believe to be the proper reading frame and the predicted amino acid sequence of the DH1308_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:18.
3 0 - The EcoRI/NotI restriction fragment obtainable from the deposit containing clone DH1308_1 should be approximately 3300 bp.
The nucleotide sequence disclosed herein for DH1308_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DH1308_1 demonstrated at least some similarity with sequences identified as W13942 (mb23h02.r1 Soares mouse p3NMF19.5 Mus musculus cDNA
clone 3302915'). Based upon sequence similarity, DH1308_1 proteins and each similar protein or peptide may share at least some activity.
Clone "DL185 1"
A polynucleotide of the present invention has been identified as clone "DL185 1".
DL185_1 was isolated from a human adult brain cDNA library using methods which are selective for cDNAs encoding secreted proteins (see U.S. Pat. No. 5,536,637), or was identified as encoding a secreted or transmembrane protein on the basis of computer analysis of the amino acid sequence of the encoded protein. DL185_1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "DL185_1 protein').
The nucleotide sequence of DL185_1 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 DL185_1 protein corresponding to the foregoing nucleotide sequence is reported in SEQ ID N0:20. Amino acids 7 to 19 are a predicted leader/signal sequence, with the predicted mature amino acid sequence beginning at amino acid 20, or are a transmembrane domain.
The EcoRI/NotI restriction fragment obtainable from the deposit containing clone 2 0 DL185_1 should be approximately 3480 bp.
The nucleotide sequence disclosed herein for DL185_1 was searched against the GenBank and GeneSeq nucleotide sequence databases using BLASTN/BLASTX and FASTA search protocols. DL185_1 demonstrated at least some similarity with sequences identified as D87248 (Rat mRNA for NB-3, complete cds), U35371 (Rattus norvegicus 2 5 neural cell adhesion protein BIG-2 precursor (BIG-2) mRNA, complete cds), and X99043 (M.musculus mRNA for brain-derived immunoglobulin superfamiiy molecule BIG-2).
The predicted amino acid sequence disclosed herein for DL185_1 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The predicted DL185_1 protein demonstrated at least some similarity to 3 0 sequences identified as D87248 {NB-3 [Rattus norvegicus]), L01991 (neuronal glycoprotein [Mus musculus]), R63759 (Human contactin (EMBL Accession #Z21488)), and X99043 (brain-derived immunoglobulin superfamily molecule [Mus musculus]). Contactin proteins mediate cell surface interactions during nervous system development and are attached to the membrane by a glycosylphosphatidylinositol anchor. Based upon wo ~s~os9s rcr~s9sioosa3 sequence similarity, DL185_1 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 DL185_1 protein sequence centered around amino acid 1010 of SEQ ID N0:20. The DL185_1 protein sequence also contains an ABC
transporter motif at amino acid 639 of SEQ ID N0:20. ABC transporters are involved in an extracellular transport pathway for proteins that lack signal sequences.
Devosit of Clones Clones BH272_3, BP202_3, CH27 1, CI542_2, CN483_2, C01224 2, CW768_1, CZ268_l, DH1308_1 and DL185_1 were deposited on January 8,1997 with the American Type Culture Collection as an original deposit under the Budapest Treaty and were given the accession number ATCC 98289, from which each clone comprising a particular polynucleotide is obtainable. All restrictions on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, except for the requirements specified in 37 C.F.R. ~ 1.808(b).
Each clone has been transfected into separate bacterial cells (E. coli) in this composite deposit. Each clone can be removed from the vector in which it was deposited by performing an EcoRI/NotI digestion (5' site, EcoRI; 3' site, NotI) to produce the appropriate fragment for such clone. Each clone was deposited in either the pED6 or 2 0 pNOTs vector depicted in Fig. 1. The pED6dpc2 vector ("pED6") was derived from pED6dpcl by insertion of a new polylinker to facilitate cDNA cloning (Kaufman et al., 1991, Nucleic Acids Res. 19: 4485-4490); the pNOTs vector was derived from pMT2 (Kaufman et al., 1989, Mol. Cell. Biol. 9: 946-958) by deletion of the DHFR
sequences, insertion of a new polylinker, and insertion of the M13 origin of replication in the CIaI site.
2 5 In some instances, the deposited clone can become "flipped" (i.e., in the reverse orientation) in the deposited isolate. In such instances, the cDNA insert can still be isolated by digestion with EcoRI and NotI. However, NotI will then produce the 5' site and EcoRI will produce the 3' site for placement of the cDNA in proper orientation for expression in a suitable vector. The cDNA may also be expressed from the vectors in 3 0 which they were deposited.
Bacterial cells containing a particular clone can be obtained from the composite deposit as follows:
An oligonucleotide probe or probes should be designed to the sequence that is known for that particular clone. This sequence can be derived from the sequences provided herein, or from a combination of those sequences. The sequence of the oligonucleotide probe that was used to isolate each full-length clone is identified below, and should be most reliable in isolating the clone of interest.
Clone Probe Sequence BH272_3 SEQ ID N0:21 BP202_3 SEQ ID N0:22 CH27 1 S EQ ID N0:23 CI542_2 SEQ ID N0:24 CN483_2 SEQ ID N0:25 C01224_2 SEQ ID N0:26 CW768_1 SEQ ID N0:27 CZ268_1 SEQ ID N0:28 DH1308_1 SEQ ID N0:29 DL185_1 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 (1-2 0 dimethoxytrityloxy-2-(N-biotinyl-4-aminobutyl)-propyl-3-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramadite) (Glen Research, cat. no. 10-1953)).
The design of the oligonucleotide probe should preferably follow these parameters:
(a) It should be designed to an area of the sequence which has the fewest 2 5 ambiguous bases ("N's"), if any;
(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-3~P ATP {specific activity 6000 Ci / mmole) and T4 polynucleotide lcinase using commonly employed techniques for 3 0 labeling oligonucleotides. Other labeling techniques can also be used.
Unincorporated label should preferably be removed by gel filtration chromatography or other established methods. The amount of radioactivity incorporated into the probe should be quantitated by measurement in a scintillation counter. Preferably, specific activity of the resulting probe should be approximately 4e+6 dpm/pmole.

wo 9sr3o69s rc1'rtrs9sroosa3 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 pg/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 lzg/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, I00 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 1 a+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 1~,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 earner 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 wo 9sr3o69s rcT~rs9sioosa3 the Fc portion of an immunoglobulin. For a bivalent form of the protein, such a fusion could be to the Fc portion of an IgG molecule. Other immunoglobulin isotypes may also be used to generate such fusions. For example, a protein - IgM fusion would generate a decavalent form of the protein of the invention.
The present invention also provides both full-length and mature forms of the disclosed proteins. The full-length form of the such proteins is identified in the sequence listing by translation of the nucleotide sequence of each disclosed clone. The mature form of such protein may be obtained by expression of the disclosed full-length polynucleotide (preferably those deposited with ATCC) in a suitable mammalian cell or other host cell.
The sequence of the mature form of the protein may also be determinable from the amino acid sequence of the full-length form.
The present invention also provides genes corresponding to the polynudeotide 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, enhanrers, 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 sourres 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 mlZIVA transcribed from the gene (Albert and Morris,1994, Trends Pharmacol. Sci.15(7): 250-254; Lavarosky et aL, 1997, 3 0 Biochem. MoI. 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 Bl, 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 WO 98J30695 PCT/US98ro0543 shares at least 75% sequence identity (more preferably, at least 85% identity;
most preferably at least 95% identity) with any such segment of any of the disclosed proteins.
Species homologs of the disclosed polynucleotides and proteins are also provided by the present invention. As used herein, a "species homologue" is a protein or polynucleotide with a different species of origin from that of a given protein or polynucleotide, but with significant sequence similarity to the given protein or polynucleotide, as determined by those of skill in the art. Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source from the desired species.
The invention also encompasses allelic variants of the disclosed polynucleotides or proteins; that is, naturally-occurring alternative forms of the isolated polynucleotide which also encode proteins which are identical, homologous, or related to that encoded by the polynucleotides .
The invention also includes polynucleotides with sequences complementary to those of the polynucleotides disclosed herein.
The present invention also includes polynucleotides capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency conditions are shown in the table below: highly stringent conditions are those that are at 2 0 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.

WO 98/30695 PC~'IUS98~00543 StringencyPolynucleotideHybridHybridization TemperatureWash ConditionHybrid Lengthand Temperature ~P)# Buffer' and Buffers A DNA:DNA s 50 65C; lxSSC -or- 65C; 0.3xSSC
42C; lxSSC, 50% formamide B DNA:DNA <50 TB*; lxSSC TB*; lxSSC

C DNA:RNA 2 50 67C; lxSSC -or- 67C; 0.3xSSC
45C; lxSSC, 50% formamide D DNA:RNA <50 TD*; lxSSC Tp*; ixSSC

E RNA:RNA 2 50 70C; lxSSC -or- 70C; 0.3xSSC
50C; lxSSC, 50% formamide F RNA:RNA <50 TF"; lxSSC TF*; lxSSC

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

I DNA:RNA 2 50 67C; 4xSSC -or- 67C; lxSSC
45C; 4xSSC, 50% formamide J DNA:RNA <50 T~*; 4xSSC T~*; 4~

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

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

O DNA:RNA z 50 55C; 4xSSC -or- 55C; 2xSSC
42C; 6xSSC, 50% formamide P DNA:RNA <50 Tp*; 6xSSC Tp*; 6xSSC

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

x: The hybrid length is that anticipated for the hybridized regions) of the hybridizing polynudeotides. 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.
': 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 l5mM sodium citrate) in the hybridization and wash buffers; washes are performed for 15 minutes 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 lxSSC = 0.165 M).

wo 9s~s9s rows Additional examples of stringency conditions for polynucleotide hybridization are provided in Sambrook, J., E.F. Fritsch, and T. Maruatis, 1989, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Laboratory Press, Coid Spring Harbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology,1995, F.M.
Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and 6.3-6.4, incorporated herein by reference.
Preferably, each such hybridizing polynucleotide has a length that is at least 25%(more preferably at least 50%, and most preferably at least 75%) of the length of the polynucleotide of the present invention to which it hybridizes, and has at least 60%
sequence identity (more preferably, at least 75% identity; most preferably at least 90% or 95% identity) with the polynucleotide of the present invention to which it hybridizes, where sequence identity is determined by comparing the sequences of the hybridizing polynucleotides when aligned so as to maximize overlap and identity while minimizing sequence gaps.
The isolated polynucleotide of the invention may be operably Linked to an expression control sequence such as the pMT2 or pED expression vectors disclosed in Kaufman et al., Nucleic Acids Res. 19, 4485-4490 (1991), in order to produce the protein recombinantly. Many suitable expression control sequences are known in the art. General methods of expressing recombinant proteins are also known and are exemplified in R.
Kaufman, Methods in Enzymology 185, 537-566 (1990). As defined herein "operably 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 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., lnvitrogen, San Diego, California, U.S.A. (the MaxBacO kit), and such methods are well known in the art, as described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (198, incorporated herein by reference. As used herein, an insect cell capable of expressing a polynucleotide of the present invention is "transformed."
The protein of the invention may be prepared by culturing transformed host cells under culture conditions suitable to express the recombinant protein. The resulting expressed protein may then be purified from such culture (i.e., from culture medium or cell extracts) using known purification processes, such as gel filtration and ion exchange chromatography. The purification of the protein may also include an affinity column 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 (T1ZX). 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 (1ZP-HPLC) steps employing hydrophobic ltP-HPLC media, e.g., silica gel having pendant WO 98/30695 PCT/US98~0543 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.

WO 98130695 PCT/US98ro0543 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 potentially 2 5 binds to another protein (such as, for example, in a receptor-ligand interaction), the polynucleotide can also be used in interaction trap assays (such as, for example, that described in Gyuris et al., Cell 75:791-803 (1993)) to identify polynudeotides encoding the other protein with which binding occurs or to identify inhibitors of the binding interaction.
3 0 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 wo 9~os9s rc'r~rs9sroos43 the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state); and, of course, to isolate correlative receptors or ligands. Where the protein binds or potentially binds to another protein (such as, for example, in a receptor-ligand interaction), the protein can be used to identify the other protein with which binding occurs or to identify inhibitors of the binding interaction. Proteins involved in these binding interactions can also be used to screen for peptide or small molecule inhibitors or agonists of the binding interaction.
Any or all of these research utilities are capable of being developed into reagent grade or kit format for commercialization as research products.
Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include without limitation "Molecular Cloning: A Laboratory Manual", 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E.F. Fritsch and T. Maniatis eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning Techniques", Academic Press, Berger, S.L. and A.R. Kimmel eds.,1987.
Nutritional Uses Polynucleotides and proteins of the present invention can also be used as nutritional sources or supplements. Such uses include without limitation use as a protein 2 0 or amino acid supplement, use as a carbon source, use as a nitrogen source and use as a source of carbohydrate. In such cases the protein or polynucleotide of the invention can be added to the feed of a particular organism or can be administered as a separate solid or liquid preparation, such as in the form of powder, pills, solutions, suspensions or capsules. In the case of microorganisms, the protein or polynucleotide of the invention 2 5 can be added to the medium in or on which the microorganism is cultured.
CYtokine and Cell Proliferation/Differentiation Activitx A protein of the present invention may exhibit cytokine, cell proliferation (either inducing or inhibiting) or cell differentiation (either inducing or inhibiting} activity or may 3 0 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 wo 9s rc~rrtrs~roosa3 for cell lines including, without limitation, 32D, DA2, DA1G, T10, B9, B9/I1, 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 nod a 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 Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toronto.1994.
2 0 Assays for proliferation and differentiation of hematopoietic and lymphopoietic cells include, without limitation, those described in: Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottomly, K., Davis, L.S. and Lipsky, P.E. In Current Protocols in Immunology. J.E.e.a. Coligan eds. Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toronto. 1991; deVries et al., J. Exp. Med. 173:1205-1211, 1991; Moreau et al., Nature 2 5 336:690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci. U.S.A.
80:2931-2938, 1983;
Measurement of mouse and human interleukin 6 - Nordan, R. In Current Protocols in Immunology. j.E.e.a. Coligan eds. Vol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto.1991;
Smith et al., Proc. Natl. Acad. Sci. U.S.A. 83:1857-1861, 1986; Measurement of human Interleukin 11- Bennett, F., Giannotti, J., Clark, S.C. and Turner, K. J. In Current Protocols 3 0 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.

WO 98!30695 PCT/US98~OU543 Assays for T-cell clone responses to antigens (which will identify, among others, proteins that affect APC-T cell interactions as well as direct T-cell effects by measuring proliferation and cytokine production) include, without limitation, those described in:
Current Protocols in Immunology, Ed by J. E. Coligan, A.M. Kruisbeek, D.H.
Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function; Chapter 6, Cytokines and their cellular receptors; Chapter 7, Immunologic studies in Humans);
Weinberger et al., Proc. Natl. Acad. Sci. USA 77:6091-6095, 1980; Weinberger et al., Eur. J.
Immun.

11:405-411, 1981; Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol.
140:508-512, 1988.
Immune Stimulating or Suppressing Activity A protein of the present invention may also exhibit immune stimulating or immune suppressing activity, including without limitation the activities for which assays are described herein. A protein may be useful in the treatment of various immune deficiencies and disorders (including severe combined immunodeficiency (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, herpesvinuses, 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 wo 9sr3os9s rc r~s~~oosa3 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.
l.mmunosuppression 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 rells, followed by an immune reaction that destroys the transplant. The administration of a molecule which inhibits or blocks interaction of a B7 lymphocyte antigen with its natural ligand(s) on immune cells (such as a soluble, monomeric form of a peptide having B7-2 activity alone or in conjunction with a 2 5 monomeric form of a peptide having an activity of another B lymphocyte antigen (e.g., B7-1, B7-3) or blocking antibody), prior to transplantation can lead to the binding of the molecule to the natural ligand(s) on the immune cells without transmitting the corresponding costimulatory signal. Blocking B lymphocyte antigen function in this matter prevents cytokine synthesis by immune cells, such as T cells, and thus acts as an 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. Natl. 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 wo 9sr~o69s rcr~s9s<oosa~
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 Vii=
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.
44.

WO 98/'30695 PCTIUS981~00543 Expression of the appropriate class I or class II MHC in conjunction with a peptide having the activity of a B lymphocyte antigen (e.g., B7-1, B7-2, B7-3) induces a T
cell mediated immune response against the transfected tumor cell. Optionally, a gene encoding an antisense construct which blocks expression of an MHC class II associated protein, such as the invariant chain, can also be cotransfected with a DNA encoding a peptide having the activity of a B lymphocyte antigen to promote presentation of tumor associated antigens and induce tumor specific immunity. Thus, the induction of a T cell mediated immune response in a human subject may be sufficient to overcome tumor-specific tolerance in the subject.
The activity of a protein of the invention may, among other means, be measured by the following methods:
Suitable assays for thymocyte or splenocyte cytotoxicity include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E.
Coligan, A.M.
Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Herrmann et al., Proc. Natl.
Acad. Sci.
USA 78:2488-2492,1981; Herrmann et al., J. Immunol.128:1968-1974,1982; Handa et al., J. Immunol.135:1564-1572,1985; Takai et al., J. Immunol.137:3494-3500,1986;
Takai et al., J. Immunol.140:508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci. USA
78:2488-2492, 2 0 1981; Herrmann et al., J. Immunol. 128:1968-1974, 1982; Handa et al., J.
Immunol.
135:1564-1572, 1985; Takai et al., J. Immunol. 137:3494-3500, 1986; Bowmanet al., J.
Virology 61:1992-1998; Takai et al., J. Immunol. 140:508-512, 1988;
Bertagnolli et al., Cellular Immunology 133:327-341,1991; Brown et al., J. Immunol. 153:3079-3092, 1994.
Assays for T-cell-dependent immunoglobulin responses and isotype switching 2 5 (which will identify, among others, proteins that modulate T-cell dependent antibody responses and that affect Th1 /Th2 profiles) include, without limitation, those described in: Maliszewski, J. Immunol. 144:3028-3033, 1990; and Assays for B cell function: In vitro antibody production, Mond, J.J. and Brunswick, M. In Current Protocols in Immunology.
J.E.e.a. Coligan eds. Vol 1 pp. 3.8.1-3.8.16, John Wiley and Sons, Toronto.1994.
3 0 Mixed lymphocyte reaction (MLR) assays (which will identify, among others, proteins that generate predominantly Thl and CTL responses) include, without limitation, those described in: Current Protocols in Immunology, Ed by J. E. Coligan, A.M.
Kruisbeek, D.H. Margulies, E.M. Shevach, W Strober, Pub. Greene Publishing Associates and Wiley-Interscience (Chapter 3, In Vitro assays for Mouse Lymphocyte Function 3.1-3.19; Chapter 7, Immunologic studies in Humans); Takai et al., J. Immunol. 137:3494-3500, 1986; Takai et al., J. Immunol. 140:508-512, 1988; Bertagnolli et al., J. Immunol.
149:3778-3783,1992.
Dendritic cell-dependent assays (which will identify, among others, proteins expressed by dendritic cells that activate naive T-cells) include, without limitation, those described in: Guery et al., J. Immunol. 134:536-544, 1995; Inaba et al., Journal of Experimental Medicine 173:549-559, 1991; Macatonia et al., Journal of Immunology 154:5071-5079,1995; Porgador et al., Journal of Experimental Medicine 182:255-260,1995;
Nair et al., Journal of Virology 67:4062-4069, 1993; Huang et al., Science 264:961-965, 1994; Macatonia et al., Journal of Experimental Medicine 169:1255-1264,1989;
Bhardwaj et al., Journal of Clinical Investigation 94:797-807, 1994; and Inaba et al., Journal of Experimental Medicine 172:631-640,1990.
Assays for lymphocyte survival / apoptosis (which will identify, among others, proteins that prevent apoptosis after superantigen induction and proteins that regulate lymphocyte homeostasis) include, without limitation, those described in:
Darzynkiewicz et al., Cytometry 13:795-808,1992; Gorczyca et al., Leukemia 7:659-670, 7 993;
Gorczyca et al., Cancer Research 53:1945-1951, 1993; Itoh et al., Cell 66:233-243, 1991;
Zacharchuk, Journal of Immunology 145:4037-4045, 1990; Zamai et al., Cytometry 14:891-897, 1993;
Gorczyca et al., International Journal of Oncology 1:639-648, 1992.
Assays for proteins that influence early steps of T-cell commitment and 2 0 development include, without limitation, those described in: Antica et al., Blood 84111-117, 1994; Fine et al., Cellular Immunology 155:111-122, 1994; Galy et al., Blood 85:2770-2778,1995; Toki et al., Proc. Nat. Acad Sci. USA 88:7548-7551,1991.
Hematopoiesis Regulatin~~ Activity 2 5 A protein of the present invention may be useful in regulation of hematopoiesis and, consequently, in the treatment of myeloid or lymphoid cell deficiencies.
Even marginal biological activity in support of colony forming cells or of factor-dependent cell lines indicates involvement in regulating hematopoiesis, e.g. in supporting the growth and proliferation of erythroid progenitor cells alone or in combination with other cytokines, 3 0 thereby indicating utility, for example, in treating various anemias or for use in conjunction with irradiation/chemotherapy to stimulate the production of erythroid precursors and/or erythroid cells; in supporting the growth and proliferation of myeloid cells such as granulocytes and monocytes/macrophages (i.e., traditional CSF
activity) useful, for example, in conjunction with chemotherapy to prevent or treat consequent wo 9s rcrrvs9sioosa~
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 hematopoietfc lines are cited above.
Assays for embryonic stem cell differentiation (which will identify, among others, proteins that influence embryonic differentiation hematopoiesis) include, without limitation, those described in: Johansson et al. Cellular Biology 15:141-151,1995; Keller et 2 0 al., Molecular and Cellular Biology 13:473-486, 1993; McClanahan et al., Blood 81:2903-2915,1993.
Assays for stem cell survival and differentiation (which will identify, among others, proteins that regulate lympho-hematopoiesis) include, without limitation, those described in: Methylceliulose colony forming assays, Freshney, M.G. In Culture of 2 5 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, LK. and Briddell, R.A. In Culture of Hematopoietic Cells. R.I. Freshney, et al. eds.
Vol pp. 23-39, Wiley-Liss, lnc., New York, NY.1994; Neben et al., Experimental Hematology 22:353-359, 3 0 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 Ctdture of Hematopoietic Cells. R.I. Freshney, et al. eds. Vol pp. 163-179, Wiley-Liss, lnc., New York, NY.1994; Long term culture initiating cell assay, Sutherland, WO 9$/30695 PCT/US98/00543 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 bums, incisions and ulcers.
A protein of the present invention, which induces cartilage and / or bone growth in circumstances where bone is not normally formed, has application in the healing of bone fractures and cartilage damage or defects in humans and other animals.
Such a preparation employing a protein of the invention may have prophylactic use in closed as well as open fracture reduction and also in the improved fixation of artificial joints. De novo bone formation induced by an osteogenic agent contributes to the repair of congenital, trauma induced, or oncologic resection induced craniofacial defects, and also is useful in cosmetic plastic surgery.
A protein of this invention may also be used in the treatment of periodontal disease, and in other tooth repair processes. Such agents may provide an environment to attract bone-forming cells, stimulate growth of bone-forming cells or induce 2 0 differentiation of progenitors of bone-forming cells. A protein of the invention may also be useful in the treatment of osteoporosis or osteoarthritis, such as through stimulation of bone and/or cartilage repair or by blocking inflammation or processes of tissue destruction (collagenase activity, osteoclast activity, etc.) mediated by inflammatory processes.
2 5 Another category of tissue regeneration activity that may be attributable to the protein of the present invention is tendon/ligament formation. A protein of the present invention, which induces tendon/ligament-like tissue or other tissue formation in circumstances where such tissue is not normally formed, has application in the healing of tendon or ligament tears, deformities and other tendon or ligament defects in humans and 3 0 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 rells 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, I-iuntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome. Further conditions which may be treated in accordance with the present 2 0 invention include mechanical and traumatic disorders, such as spinal cord disorders, head trauma and cerebrovascular diseases such as stroke. Peripheral neuropathies resulting from chemotherapy or other medical therapies may also be treatable using a protein of the invention.
Proteins of the invention may also be useful to promote better or faster closure of 2 5 non-healing wounds, including without limitation pressure ulrers, ulcers associated with vascular insufficiency, surgical and traumatic wounds, and the like.
It is expected that a protein of the present invention may also exhibit activity for generation or regeneration of other tissues, such as organs (including, for example, pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac) 3 0 and vascular (including vascular endothelium) tissue, or for promoting the growth of cells comprising such tissues. Part of the desired effects may be by inhibition or modulation of fibrotic scarring to allow normal tissue to regenerate. A protein of the invention may also exhibit angiogenic activity.

wo 9sr~s9s rcTivs9sioosa3 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, Inc., Chicago, as modified by Eaglstein and Mertz, J.
Invest.
Dermatol 71:382-84 (1978).
Activin/Inhibin Activity A protein of the present invention may also exhibit activin- or inhibin-related 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-(3 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:

wo s p~,US
Assays for activin/inhibin activity include, without limitation, those described in:
Vale et al., Endocrinology 91:562-572, 1972; Ling et al., Nature 321:779-782,1986; Vale et al., Nature 321:776-779,1986; Mason et al., Nature 318:659-663,1985; Forage et al., Proc.
Natl. Acad. Sci. USA 83:3091-3095,1986.
Chemotactic/Chemokinetic Activity A protein of the present invention may have chemotactic or chemokinetic activity (e.g., act as a chemokine) for mammalian cells, including, for example, monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and /
or endothelial rells.
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 indure 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. ICruisbeek, D.H. Margulies, E.M. Shevach, W.Strober, Pub.
Greene 3 0 Publishing Associates and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chemokines 6.12.1-6.12.28; Taub et al. J. Clin. Invest. 95:1370-1376,1995; Lind et al.
APMIS 103:140-146, 1995; Muller et al Eur. J. Immunol. 25:1744-1748; Gruber et al. j. of Immunol. 152:5860-5867,1994; Johnston et al. J. of Immunol.153:1762-1768,1994.

Hemostatic and Thrombol3rtic 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 Iigands) may themselves be useful as inhibitors of receptor/Iigand 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-Inflammatory Activity Proteins of the present invention may also exhibit anti-inflammatory activity.
The anti-inflammatory activity may be achieved by providing a stimulus to cells involved in the inflammatory response, by inhibiting or promoting cell-cell interactions (such as, for example, cell adhesion), by inhibiting or promoting chemotaxis of cells involved in the inflammatory process, inhibiting or promoting cell extravasaHon, 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 Suppressor Activity Cadherins are calcium-dependent adhesion molecules that appear to play major 2 5 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 wo ~os~s rcrr~rs~osa~
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 earner) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient(s). The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the invention may also contain cytokines, lymphokines, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IF1V, TNFO, TNFl, T1VF2, G-CSF, Meg-CSF, thrombopoietin, stem 2 0 cell factor, and erythropoietin. The pharmaceutical composition may further contain other agents which either enhance the activity of the protein or compliment its activity or use in treatment. Such additional factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with protein of the invention, or to minimize side effects. Conversely, protein of the present invention may be included 2 5 in formulations of the particular cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent to minimize side effects of the cytokine, lymphokine, other hematopoietic factor, thrombolytic or anti-thrombotic factor, or anti-inflammatory agent.
A protein of the present invention may be active in multimers {e. g., heterodimers 3 0 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 wo s rc~ricrs~sroos4s 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 acreptable Garners, with amphipathic agents such as lipids which exist in aggregated form as micelles, insoluble monolayers, liquid crystals, or lamellar layers in aqueous solution. Suitable lipids for liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponin, bile acids, and the like. Preparation of such liposomal formulations is within the level of skill in the art, as disclosed, for example, in U.S. Patent No. 4,235,871; U.S. Patent No.
4,501,728; U.S.
2 0 Patent No. 4,837,028; and U.S. Patent No. 4,737,323, all of which are incorporated herein by reference.
As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to show a meaningful patient benefit, i.e., treatment, healing, prevention or 2 5 amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.
3 0 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 wo 9s rcTrtrs9sioosa3 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 earner such as a gelatin or an adjuvant. The tablet, capsule, and powder contain from about 5 to 95°~o protein of the present invention, and preferably from about 25 to 90% protein of the present invention.
2 0 When administered in liquid form, a liquid earner such as water, petroleum, oils of animal or plant origin such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid 2 5 form, the pharmaceutical composition contains from about 0.5 to 90% by weight of protein of the present invention, and preferably from about 1 to 50% protein of the present invention.
When a therapeutically effective amount of protein of the present invention is administered by intravenous, cutaneous or subcutaneous injection, protein of the present 3 0 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 WO 98130695 PCT/USt98100543 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 pharmareutical 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 pg to about 100 mg (preferably about O.lng to about 10 mg, more preferably about 0.1 Ilg to about 1 mg) of protein of the present invention per kg body weight.
The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the condition and potential idiosyncratic response of each individual patient.
It is 2 0 contemplated that the duration of each application of the protein of the present invention will be in the range of 12 to 24 hours of continuous intravenous administration.
Ultimately the attending physician will decide on the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention.
Protein of the invention may also be used to immunize animals to obtain 2 5 polyclonal and monoclonal antibodies which specifically react with the protein. Such antibodies may be obtained using either the entire protein or fragments thereof as an immunogen. The peptide immunogens additionally may contain a cysteine residue at the carboxyl terminus, and are conjugated to a hapten such as keyhole limpet hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, as in 3 0 R.P. Merrifield, J. Amer.Chem.Soc. 85 2149-2154 (1963); J.L. Krstenansky, et al., FEES Lett.
211, 10 (1987). Monoclonal antibodies binding to the protein of the invention may be useful diagnostic agents for the immunodetection of the protein. Neutralizing monoclonal antibodies binding to the protein may also be useful therapeutics for both conditions associated with the protein and also in the treatment of some forms of cancer where abnormal expression of the protein is involved. In the case of cancerous cells or leukemic cells, neutralizing monoclonal antibodies against the protein may be useful in detecting and preventing the metastatic spread of the cancerous cells, which may be mediated by the protein.
For compositions of the present invention which are useful for bone, cartilage, tendon or ligament regeneration, the therapeutic method includes administering the composition topically, systematically, or locally as an implant or device.
When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of bone, cartilage or tissue damage. Topical administration may be suitable for wound healing and tissue repair. Therapeutically useful agents other than a protein of the invention which may also optionally be included in the composition as described above, may alternatively or additionally, be administered simultaneously or sequentially with the composition in the methods of the invention. Preferably for bone and / or cartilage formation, the composition would include a matrix capable of delivering the protein-containing composition to the site of bone and/or cartilage damage, providing a structure for the developing bone and cartilage and optimally capable of being resorbed into the body.
Such matrices may be formed of materials presently in use for other implanted medical 2 0 applications.
The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions will define the appropriate formulation.
Potential matrices for the compositions may be biodegradable and chemically defined calcium 2 5 sulfate, tricalciumphosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydrides. Other potential materials are biodegradable and biologically well-defined, such as bone or dermal collagen. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically defined, such as sintered hydroxapatite, bioglass, aluminates, or other 3 0 ceramics. Matrices may be comprised of combinations of any of the above mentioned types of material, such as polylactic acid and hydroxyapaHte 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.

wo s rcrms98~oosa3 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.
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 2 0 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-(3), and insulin-like growth factor (IGF).
The therapeutic compositions are also presently valuable for veterinary applications. Particularly domestic animals and thoroughbred horses, in addition to 2 5 humans, are desired patients for such treatment with proteins of the present invention.
The dosage regimen of a protein-containing pharmaceutical composition to be used in tissue regeneration will be determined by the attending physician considering various factors which modify the action of the proteins, e.g., amount of tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of 3 0 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 WO 98130695 PCT/US98~0543 the dosage. Progress can be monitored by periodic assessment of tissue/bone growth and/or repair, for example, X-rays, histomorphometric determinations and tetracycline labeling.
Polynucleotides of the present invention can also be used for gene therapy.
Such polynucleotides can be introduced either in vivo or ex vivo into cells for expression in a mammalian subject. Polynucleotides of the invention may also be administered by other known methods for introduction of nucleic acid into a cell or organism (including, without limitation, in the form of viral vectors or naked DNA).
Cells may also be cultured ex vivo in the presence of proteins of the present invention in order to proliferate or to produce a desired effect on or activity in such cells.
Treated cells can then be introduced in vivo for therapeutic purposes.
Patent and literature references cited herein are incorporated by reference as if fully set forth.

SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Jacobs, Kenneth McCoy, John M.
LaVallie, Edward R.
Racie, Lisa A.
Merberg, David Treacy, Maurice Spaulding, Vikki Agostino, Michael J.
(ii) TITLE OF INVENTION: SECRETED PROTEINS AND POLYNUCLEOTIDES
ENCODING THEM
(iii) NUMBER OF SEQUENCES: 30 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Genetics Institute, Inc.
(B) STREET: 87 CambridgePark Drive (C) CITY: Cambridge (D) STATE: MA
(E) COUNTRY: U.S.A.
(F) ZIP: 02140 (v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0) Version #1.30 (vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Sprunger, Suzanne A.
(B) REGISTRATION NUMBER: 41,323 (ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: (617) 498-8284 (B) TELEFAX: (617) 876-5851 (2) INFORMATION FOR SEQ ID N0:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 886 base pairs (B) TYpE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear wo ~s~os9s rc~rms9sroosa~
(ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION:
SEQ ID N0:1:

GTAATAATAA AGAAATTTCT AACAAAAAAAF,~~.~.AAAAAAA AP~AAAA 8 (2) INFORMATION FOR SEQ
ID N0:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 178 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2:
Met Pro Ala Tyr His Ser Ser Leu Met Asp Pro Asp Thr Lys Leu Ile Gly Asn Met AlaLeu Leu IleArg Ser Gln LysGly Pro Ala Pro Phe Pro Arg Glu ThrLys Asp AspIle Val Asp AlaIle Tyr Tyr Thr Glu Phe Lys Ala AsnVal Phe LysAsn Tyr Glu LysAsn Glu Ala Phe Ile Asp Arg Thr Leu Ile Tyr Ile Thr Leu Tyr Ile Ser Glu Cys Leu Lys Lys Leu Gln Lys Cys Asn Ser Lys Ser Gln Gly Glu Lys Glu Met Tyr Thr Leu Gly Ile Thr Asn Phe Pro Ile Pro Gly Glu Pro Gly Phe Pro Leu Asn Ala Ile Tyr Ala Lys Pro Ala Asn Lys Gln Glu Asp Glu Val Met Arg Ala Tyr Leu Gln Gln Leu Arg Gln Glu Thr Gly Leu Arg Leu Cys Glu Lys Val Phe Asp Pro Gln Asn Asp Lys Pro Ser Lys Trp Trp Thr Cys Phe Val Lys Arg Gln Phe Met Asn Lys Ser Leu Ser Gly Pro Gly Gln {2) INFORMATION FOR SEQ ID N0:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1537 base pairs (B) TYPE: nucleic acid (C)'STRANDEDNESS: double (D) TOPOLOGY: linear {ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:3:

AGTAATATTC

AAACAAAAGA

GAGTTTCCAA

TTACCTGGTT

GCTTTTGCAT

GGTTCTCGAT

TCCCAGAAAC

GGTGATGCTG

AAGCAACAGA

AGTACAACAC

GATGATGATA

CCATTGGTAA

AGGAATGACT

ATTGTCTGGG

AGAAGGAAAA

ACTAATGCTG

TCTGCATAGT

TTGTATTTTC

GACTTTTTTG

TACTAAAAAT

ATATTAAAAT

ATATATAGAT AATTTTTTAA 1?~~AAAAAAAA AAAAAAA 1537 (2) INFORMATION FOR SEQ ID N0:4:

{i} SEQUENCE CHARACTERISTICS:

(A) LENGTH: 397 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein wo 9s rc~r~s~sroos43 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4:
Met Leu Thr Ser Asp Phe Lys Lys Asn Thr Arg Leu Leu Pro Lys Leu Lys Lys Ile Glu Asn Gln Val Ala Met Ser Phe Tyr Lys His Gln Ser Ser Pro Asp Leu Ser Ser Glu Glu Ser Glu Thr Glu Lys Glu Ile Lys Arg Lys Ala Glu Val Lys Lys Thr Lys Ala Gly Asn Thr Lys Glu Ala Val Val His Leu Arg Lys Ser Thr Arg Asn Thr Ser Asn Ile Pro Val Ile Leu Glu Pro Glu Thr Glu Glu Ser Glu Asn Glu Phe Tyr Ile Lys Gln Lys Lys Ala Arg Pro Ser Val Lys Glu Thr Leu Gln Lys Ser Gly Val Arg Lys Glu Phe Pro Ile Thr Glu Ala Val Gly Ser Asp Lys Thr Asn Arg His Pro Leu Glu Cys Leu Pro Gly Leu Ile Gln Asp Lys Glu Trp Asn Glu Lys Glu Leu Gln Lys Leu His Cys Ala Phe Ala Ser Leu Pro Lys His Lys Pro Gly Phe Trp Ser Giu Val Ala Ala Ala Val Gly Ser Arg Ser Pro Glu Glu Cys Gln Arg Lys Tyr Met Glu Asn Pro Arg Gly Lys Gly Ser Gln Lys His Val Thr Lys Lys Lys Pro Ala Asn Ser Lys Gly Gln Asn Gly Lys Arg Gly Asp Ala Asp Gln Lys Gln Thr Ile Lys Ile Thr Ala Lys Val Gly Thr Leu Lys Arg Lys Gln Gln Met Arg Glu Phe Leu Glu Gln Leu Pro Lys Asp Asp His Asp Asp Phe Phe Ser Thr Thr Pro Leu Gln His Gln Arg Ile Leu Leu Pro Ser Phe Gln Asp Ser Glu Asp Asp Asp Asp Ile Leu Pro Asn Met Asp Lys Asn Pro Thr Thr Pro Ser Ser Val Ile Phe Pro Leu Val Lys Thr Pro Gln Cys Gln His Val Ser Pro Gly Met Leu Gly Ser Ile Asn Arg Asn Asp Cys Asp Lys Tyr Val Phe Arg Met Gln Lys Tyr His Lys Ser Asn Gly Gly Ile Val Trp Gly Asn Ile Lys Lys Lys Leu Val Glu Thr Asp Phe Ser Thr Pro Thr Pro Arg Arg Lys Thr Pro Phe Asn Thr Asp Leu Gly Glu Asn Ser Gly Ile Gly Lys Leu Phe Thr Asn Ala Val Glu Ser Leu Asp Glu Glu Glu Lys Asp Tyr Tyr Phe Ser Asn Ser Asp Ser Ala (2) INFORMATION FOR SEQ ID N0:5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 921 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:5:

GTGGAGATGT TTTTGTCTTG TCCAAATAAA AGATTCACCA GTI?,AAAAp,AA AAAAAAAAAA 840 AAA,AAAAAAA AAAAAAAAAA ~1,~~~AAAAAAA p,~~i~AAAAp,AA AAAAAAAAAA AAAAAAAAAA 9 0 P~~i~AAAAAAA p,~~e~AAAAAAA A 921 (2) INFORMATION FOR SEQ ID N0:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 192 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:6:
Met Val Arg Ala Gly Ala Val Gly Ala His Leu Pro Ala Ser Gly Leu Asp Ile Phe Gly Asp Leu Lys Lys Met Asn Lys Arg Gln Leu Tyr Tyr Gln Val Leu Asn Phe Ala Met Ile Val Ser Ser Ala Leu Met Ile Trp Lys Gly'Leu Ile Val Leu Thr Gly Ser Glu Ser Pro Ile Val Val Val Leu Ser Gly Ser Met Glu Pro Ala Phe His Arg Gly Asp Leu Leu Phe Leu Thr Asn Phe Arg Glu Asp Pro Ile Arg Ala Gly Glu Ile Val Val Phe Lys Val Glu Gly Arg Asp Ile Pro Ile Val His Arg Val Ile Lys Val His Glu Lys Asp Asn Gly Asp Ile Lys Phe Leu Thr Lys Gly Asp Asn Asn Glu Val Asp Asp Arg Gly Leu Tyr Lys Glu Gly Gln Asn Trp wo ~s~os9s rc~rrt~s9sroosa~
Leu Glu Lys Lys Asp Val Val Gly Arg Ala Arg Gly Phe Leu Pro Tyr Val Gly Met Val Thr Ile Ile Met Asn Asp Tyr Pro Lys Phe Lys Tyr Ala Leu Leu Ala Val Met Gly Ala Tyr Val Leu Leu Lys Arg Glu Ser (2) INFORMATION FOR SEQ ID N0:7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1505 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:7:

ARCCGCCCCC

TCCGCGCCCG

TACGCCTTTG

CGCGACTTCA

CGGGGCCCGG

GGCGCGCTGT

GAAAACAGTG

GAAGATGGAG

CCTGAGGGGC

CTAGAGAAGC

GAGGCCTCGC

CTGTATGAGG

CAGGAGCTGG

GGGGCACTTG

CTGCTCCTGA

WO 9$/30695 PCTIUS98/00543 TTTTTCATAA Fr,FU~,AAAAAAA AAAAAAAAAA P,~~~,t~AAAAAA AAAAAAAAAA AAAAAAAAAA 15 0 AAAAA

(2) INFORMATION FOR SEQ ID N0:8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 427 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:8:
Met Tyr Ala Phe Val Arg Phe Leu Glu Asp Asn Val Cys Tyr Ala Leu Pro Val Ser Cys Val Arg Asp Phe Ser Pro Arg Ser Arg Leu Asp Phe Asp Asn Gln Lys Val Tyr Ala Val Tyr Arg Gly Pro Glu Glu Leu Gly Ala Gly Pro Glu Ser Pro Pro Arg Ala Pro Arg Asp Trp Gly Ala Leu Leu Leu His Lys Ala Gln Ile Leu Ala Leu Ala Glu Asp Lys Ser Asp Leu Glu Asn Ser Val Met Gln Lys Lys Ile Lys Ile Pro Lys Leu Ser wo ~s~os~s rcT~rs9sroosa3 Leu Asn His Val Glu Glu Asp Gly Glu Val Lys Asp Tyr Gly Glu Glu Asp Leu Gln Leu Arg His Ile Lys Arg Pro Glu Gly Arg Lys Pro Ser Glu Val Ala His Lys Ser Ile Glu Ala Val Val Ala Arg Leu Glu Lys Gln Asn Gly Leu Ser Leu Gly His Ser Thr Cys Pro Glu Glu Val Phe Val Glu Ala Ser Pro Gly Thr Glu Asp Met Asp Ser Leu Glu Asp Ala Val Val Pro Arg Ala Leu Tyr Glu Glu Leu Leu Arg Asn Tyr Gln Gln Gln Gln Glu Glu Met Arg His Leu Gln Gln Glu Leu Glu Arg Thr Arg Arg Gln Leu Val Gln Gln Ala Lys Lys Leu Lys Glu Tyr Gly Ala Leu Val Ser Glu Met Lys Glu Leu Arg Asp Leu Asn Arg Arg Leu Gln Asp Val Leu Leu Leu Arg Leu Gly Ser Gly Pro Ala Ile Asp Leu Glu Lys Val Lys Ser Glu Cys Leu Glu Pro Glu Pro Glu Leu Arg Ser Thr Phe Ser Glu Glu Ala Asn Thr Ser Ser Tyr Tyr Pro Ala Pro Ala Pro Val Met Asp Lys Tyr Ile Leu Asp Asn Gly Lys Val His Leu Gly Ser Gly Ile Trp Val Asp Glu Glu Lys Trp His Gln Leu Gln Val Thr Gln Gly Asp Ser Lys Tyr Thr Lys Asn Leu Ala Val Met Ile Trp Gly Thr Asp Val Leu Lys Asn Arg Ser Val Thr Gly Val Ala Thr Lys Lys Lys Lys Asp Ala Val Pro Lys Pro Pro Leu Ser Pro His Lys Leu Ser Ile Val Arg Glu Cys Leu Tyr Asp Arg Ile Ala Gln Glu 'r'hr Val Asp Glu Thr Glu Ile Ala Gln Arg Leu Ser Lys Val Asn Lys Tyr Ile Cys Glu Lys wo 9w3os~s rcr~us~roos~3 Ile Met Asp Ile Asn Lys Ser Cys Lys Met Lys Asn Glu Gly Lys Gln Asn Thr Ile Cys Asn Lys Leu Trp Ile Phe His (2) INFORMATION FOR SEQ ID N0:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2033 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:9:

GCCGGCCTGG

GATGCTGTGT

TGATGACTGG

AAAATCTCAG

TGCTGATGAA

RTGTKAAAAG

CTTAAATAAG

TATGCATTAT

TCTCAATGGA

TAATTTTAAG

GGATCCATAT

TACCGAGCTG

CTTTCTGTTC

TCAGGCTGAA

GACTGGAAGT

AAAATACTAT

TGCAGTTACA

GGAATTTATT

AATTATGGCA

GAGACATATA

GACGGCAGGA

ATAGGGGCCA

GAGAGATTTT

AAGTGCTCCG

CCAGTCATAA

AAGGCACACC

AAGACACATC

AAGCCGCAGG

CACGTGGACA

GCCTCAAGTC

CAAGGCGACA

AAGCAAATTG

GTTTATTGAC

TAGCCAATAT GTCATTAAAA TTAAGGTTTA TATTGTGAAA p~~?~AAAAAAA 2033 AAA

(2) INFORMATION FOR SEQ ID N0:10:

(i} SEQUENCE CHARACTERISTICS:

(A) LENGTH: 366 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:10:
Met Leu Cys Ser Leu Leu Leu Cys Glu Cys Leu Leu Leu Val Ala Gly Tyr Ala His Asp Asp Asp Trp Ile Asp Pro Thr Asp Met Leu Asn Tyr Asp Ala Ala Ser Gly Thr Met Arg Lys Ser Gln Ala Lys Tyr Gly Ile Ser Gly Glu Lys Asp Val Ser Pro Asp Leu Ser Cys Ala Asp Glu Ile Ser Glu Cys Tyr His Lys Leu Asp Ser Leu Thr Tyr Lys Ile Asp Glu 65 70 75 gp Cys Xaa Lys Lys Lys Arg Glu Asp Tyr Glu Ser Gln Ser Asn Pro Val Phe Arg Arg Tyr Leu Asn Lys Ile Leu Ile Glu Ala Gly Lys Leu Gly Leu Pro Asp Glu Asn Lys Gly Asp Met His Tyr Asp Ala Glu Ile Ile Leu Lys Arg Glu Thr Leu Leu Glu Ile Gln Lys Phe Leu Asn Gly Glu Asp Trp Lys Pro Gly Ala Leu Asp Asp Ala Leu Ser Asp Ile Leu Ile Asn Phe Lys Phe His Asp Phe Glu Thr Trp Lys Trp Arg Phe Glu Asp Ser Phe Gly Val Asp Pro Tyr Asn Val Leu Met Val Lei: Leu Cys Leu Leu Cys Ile Val Val Leu Val Ala Thr Glu Leu Trp Thr Tyr Val Arg Trp Tyr Thr Gln Leu Arg Arg Val Leu Ile Ile Ser Phe Leu Phe Ser Leu Gly Trp Asn Trp Met Tyr Leu Tyr Lys Leu Ala Phe Ala Gln His Gln Ala Glu Val Ala Lys Met Glu Pro Leu Asn Asn Val Cys Ala Lys Lys Met Asp Trp Thr Gly Ser Ile Trp Glu Trp Phe Arg Ser Ser Trp Thr Tyr Lys Asp Asp Pro Cys Gln Lys Tyr Tyr Glu Leu Leu Leu Val Asn Pro Ile Trp Leu Val Pro Pro Thr Lys Ala Leu AIa Val Thr Phe Thr Thr Phe Val Thr Glu Pro Leu Lys His Ile Gly Lys Gly Thr Gly wo ~o69s rcr~s9s~oosa3 Glu Phe Ile Lys Ala Leu Met Lys Glu Ile Pro Ala Leu Leu His Leu Pro Val Leu Ile Ile Met Ala Leu Ala Ile Leu Ser Phe Cys Tyr Gly Ala Gly Lys Ser Val His Val Leu Arg His Ile Gly Gly Pro {2) INFORMATION FOR SEQ ID N0:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2214 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:

AGAACAGAAA

ACCTTTTTTG

AGTTATCTTG

ACTATGAATT

TGTGAGAACA

CCAGATGATG

AATGATATGC

AAGACAATAG

CAGCAGAAAT

CAAGAGAAAG

TATTTTGCGC

GAGAGAGATG

AATATTAACC

GCAGACACAG

ACCAAGGAAT

GATGCTGCCT

wo 9s rc~rnls9sroos~

GAACCGGGAC CATGAGGACA GTGATCTGAT TGAAAAP,P,AC GACAGTCTGG GGAAGCGATC1500 p~~~ p~'~~ ~AAAAAAAA p~~i~.AAAAAAA ~?~~i~AAAAAAA AAAA 2 (2) INFORMATION FOR SEQ ID N0:12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 411 amino acids {B) TYPE: amino acid {C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:12:
Met Asn Ser Glu Lys Ser Met His Arg Lys Ser Thr Glu Leu Val Asn Glu Ile Thr Cys Glu Asn Thr Glu Trp Pro Gly Gln Arg Ser Thr Asn Phe Gln Ile Ile Ser Ser Tyr Pro Asp Asp Glu Ser Val Tyr Cys Thr Thr Glu Lys Tyr Asn Val Met Glu His Arg His Asn Asp Met His Tyr Glu Cys Xaa Thr Pro Cys Gln Val Thr Ser Asp Ser Asp Lys Glu Lys Thr Ile Ala Phe Leu Leu Lys Glu Leu Asp Ile Leu Arg Thr Ser Asn Lys Lys Leu Gln Gln Lys Leu Ala Lys Glu Asp Lys Glu GIn Arg Lys Leu Lys Phe Lys Leu Glu Leu Gln Glu Lys Glu Thr Glu Ala Lys Ile Ala Glu Lys Thr Ala Ala Leu Val Glu Glu Val Tyr Phe Ala Gln Lys Glu Arg Asp Glu Ala Val Met Ser Arg Leu Gln Leu Ala Ile Glu Glu Arg Asp Glu Ala Ile Ala Arg Ala Lys His Met Glu Met Ser Leu Lys Val Leu Glu Asn Ile Asn Pro Glu Glu Asn Asp Met Thr Leu Gln Glu Leu Leu Asn Arg Ile Asn Asn Ala Asp Thr Gly Ile Ala Ile Gln Lys Asn Gly Ala Ile Ile Val Asp Arg Ile Tyr Lys Thr Lys Glu Cys Lys Met Arg Ile Thr Ala Glu Glu Met Ser Ala Leu Ile GIu Glu Arg Asp Ala Ala Leu Ser Lys Cys Lys Arg Leu Glu Gln Glu Leu His His Val Lys Giu Gln Asn Gln Thr Ser Ala Asn Asn Met Arg His Leu Thr Ala Glu Asn Asn Gln Glu Arg Ala Leu Lys Ala Lys Leu Leu Ser Met Gln wo 9s rcrrt~s9sroosa~
Gln Ala Arg Glu Thr Ala Val Gln Gln Tyr Lys Lys Leu Glu Glu Glu Ile Gln Thr Leu Arg Val Tyr Tyr Ser Leu His Lys Ser Leu Ser Gln Glu Glu Asn Leu Lys Asp Gln Phe Asn Tyr Thr Leu Ser Thr Tyr Glu Glu Ala Leu Lys Asn Arg Glu Asn Ile Val Ser Ile Thr Gln Gln Gln Asn Glu Glu Leu Ala Thr Gln Leu Gln Gln Ala Leu Thr Glu Arg Ala Asn Met Glu Leu Gln Leu Gln His Ala Arg Glu Ala Ser Gln Val Ala Asn Glu Lys Val Gln Lys Leu Glu Arg Leu Val Asp Val Leu Arg Lys Lys Val Gly Thr Gly Thr Met Arg Thr Val Ile (2) INFORMATION FOR SEQ ID N0:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1227 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:13:

P~~i~AAAAAAA P,~~~AAAAAAA AAAAAAAAAG TCTGTTCTGT GGCCTCCTTC CCCCGGGCCC 1$ 0 CTGTGAGCAT

TGATGGGCCC

CTGTATAGTA

TCCTGGTCGG

CCCCCTTTCC

TAACTTCATG

CTTCAGCTCC

CTGCCAGGAG

GGATCCTCTT

ACAATGATAT

CAAAATGACA

CTTAAAGATG

(2) INFORMATION FOR SEQ ID N0:14:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 86 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:
Met Val Ile Gly Thr Asp Thr Ala Leu Cys Pro Cys Phe Leu Asp Ser Ser Ala Pro Thr Ala Asp Ser Asp Tyr Pro Leu Leu Thr Leu Leu Gly Ile Gln Thr Ala Ala Arg Arg Leu Ser Arg Tyr Leu Ser Tyr Leu Tyr Val Ile Ser Glu Arg Leu Trp Arg Ile Leu Phe Thr Trp His Pro Val Ala Ser Leu Gly Ser Gly Gly Ser Ser Asn Ser Asn Asn Asp Ile Leu 65 70 75 gp wo ~~os~ rcrrt~s His Leu Asp Asn Ser Glu (2) INFORMATION FOR SEQ ID N0:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1232 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE
DESCRIPTION:
SEQ ID
N0:15:

~1 AGTTCATTCA CAAGCATATG CTGAGAATAA ACATGTTACA CATGGGAAAA ~1~~~.AAAAAAp, 1200 p,AAAAAAAAA AAAAAAAAAA p~~ AA 12 3 2 (2) INFORMATION FOR SEQ ID N0:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 322 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:16:
Met Cys Ala Ser Tyr Ser Glu Leu Glu Leu Val Thr Ser Ala Glu Gly Leu Asn Ser Ser Gln Lys Leu Ala Cys Leu Ile Gly Val Glu Gly Gly His Ser Leu Asp Ser Ser Leu Ser Val Leu Arg Ser Phe Tyr Val Leu Gly Val Arg Tyr Leu Thr Leu Thr Phe Thr Cys Ser Thr Pro Trp Ala Glu Ser Ser Thr Lys Phe Arg His His Met Tyr Thr Asn Val Ser Gly Leu Thr Ser Phe Gly Glu Lys Val Val Glu Glu Leu Asn Arg Leu Gly Met Met Ile Asp Leu Ser Tyr Ala Ser Asp Thr Leu Ile Arg Arg Val Leu Glu Val Ser Gln Ala Pro Val Ile Phe Ser His Ser Ala Ala Arg Ala Val Cys Asp Asn Leu Leu Asn Val Pro Asp Asp Ile Leu Gln Leu Leu Lys Lys Asn Gly Gly Ile Val Met Val Thr Leu Ser Met Gly Val Leu Gln Cys Asn Leu Leu Ala Asn Val Ser Thr Val Ala Asp His Phe Asp His Ile Arg Ala Val Ile Gly Ser Glu Phe Ile Gly Ile Gly Gly WO 98J30695 PGT/US981~00543 Asn Tyr Asp Gly Thr Gly Arg Phe Pro Gln Gly Leu Glu Asp Val Ser Thr Tyr Pro Val Leu Ile Glu Glu Leu Leu Ser Arg Ser Trp Ser Glu Glu Glu Leu Gln Gly Val Leu Arg Gly Asn Leu Leu Arg Val Phe Arg Gln Val Glu Lys Val Arg Glu Glu Ser Arg Ala Gln Ser Pro VaI Glu Ala Glu Phe Pro Tyr Gly Gln Leu Ser Thr Ser Cys His Ser His Leu Val Pro Gln Asn Gly His Gln Ala Asn His Leu Glu Val Thr Lys Gln Pro Thr Asn Arg Val Pro Trp Arg Ser Ser Asn Ala Ser Pro Tyr Leu Val Ser Gly Leu Val Gly Gly Ala Thr Ile Pro Thr Phe Thr Gln Trp Phe Cys (2) INFORMATION FOR SEQ ID N0:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3320 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:17:

ACCAATGGCA

AAAGGTGGAT

GGGGTGTCAT

GTGCAGCAAG

TGTGAAAATG

CCCACACAGC

GATCTGAAAG

AGCACCCCTT

TCTTACCCCA

AATGGGAACC

AGGCTTAACG

GAGGTGCCTC

ACAAAGAGCT

CCACAAACAA

GCCAAATTGC

TCCTCTGTTG

AAAAAAGGTC

TGGTATGGCT

GCTTTGAAGT

CAGAATATTG

TATCATGTCA

ATAGCCTCTT

GAGGAGTCCA

GCTTCATCCT

CTGATGCAAG

TTTGTAAACA

AATTGGACTC

AGTTTTAAGT

wo ~os9s rcr~s~oosa3 TGCCTAAAAA p~U~Pu'4AAAAP. 3 (2) INFORMATION FOR SEQ ID N0:18:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 772 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:18:
Met Thr Ser Ser Gln Val Lys Thr Lys Pro Phe Asp Ser Trp Ser Tyr Ser Glu Met Glu Lys Glu Phe Pro Glu Leu Ile Arg Ser Val Gly Leu Leu Thr Val Ala Ala Asp Ser Ile Ser Thr Asn Gly Ser Glu Ala Val Thr Glu Glu Val Ser Gln Val Ser Leu Ser Val Asp Asp Lys Gly Gly Cys Glu Glu Asp Asn Ala Ser Ala Val Glu Glu Gln Pro Gly Leu Thr Leu Gly Val Ser Ser Ser Ser Gly Glu Ala Leu Thr Asn Ala Val Gln Pro Ser Ser Glu Thr Val Gln Gln Glu Ser Ser Ser Ser Ser His His Asp Ala Lys Asn Gln Gln Pro Val Pro Cys Glu Asn Ala Thr Pro Lys Arg Thr Ile Arg Asp Cys Phe Asn Tyr Asn Glu Asp Ser Pro Thr Gln Pro Thr Leu Pro Lys Arg Gly Leu Phe Leu Lys Glu Glu Thr Phe Lys Asn Asp Leu Lys Gly Asn Gly Gly Lys Arg Gln Met Val Asp Leu Lys Pro Glu Met Ser Arg Ser Thr Pro Ser Leu Val Asp Pro Pro Asp Arg Ser Lys Leu Cys Leu Val Leu Gln Ser Ser Tyr Pro Asn Ser Pro Ser Ala Ala Ser Gln Ser Tyr Glu Cys Leu His Lys Val Gly Asn Gly Asn Leu Glu Asn Thr Val Lys Phe His Ile Lys Glu Ile Ser Ser Ser Leu Gly Arg Leu Asn Asp Cys Tyr Lys Glu Lys Ser Arg Leu Lys Lys Pro His Lys Thr Ser Glu Glu Val Pro Pro Cys Arg Thr Pro Lys Arg Gly Thr Gly Ser Gly Lys Gln Ala Lys Asn Thr Lys Ser Ser Ala Val Pro wo 9sr~osss pcrms3 Asn Gly Glu Leu Ser Tyr Thr Ser Lys Ala Ile Glu Gly Pro Gln Thr Asn Ser Ala Ser Thr Ser Ser Leu Glu Pro Cys Asn Gln Arg Ser Trp Asn Ala Lys Leu Gln Leu Gln Ser Glu Thr Ser Ser Ser Pro Ala Phe Thr Gln Ser Ser Glu Ser Ser Val Gly Ser Asp Asn Ile Met Ser Pro Val Pro Leu Leu Ser Lys His Lys Ser Lys Lys Gly Gln Ala Ser Ser Pro Ser His Val Thr Arg Asn Gly Glu Val Val Glu Ala Trp Tyr Gly Ser Asp Glu Tyr Leu Ala Leu Pro Ser His Leu Lys Gln Thr Glu Val Leu Ala Leu Lys Leu Glu Asn Leu Thr Lys Leu Leu Pro Gln Lys Pro Arg Gly Glu Thr Ile Gln Asn Ile Asp Asp Trp Glu Leu Ser Glu Met Asn Ser Asp Ser Glu Ile Tyr Pro Thr Tyr His Val Lys Lys Lys His Thr Arg Leu Gly Arg Val Ser Pro Ser Ser Ser Ser Asp Ile Ala Ser Ser Leu Gly Glu Ser Ile Glu Ser Gly Pro Leu Ser Asp Ile Leu Ser Asp Glu Glu Ser Ser Met Pro Leu Ala Gly Met Lys Lys Tyr Ala Asp Glu Lys Ser Glu Arg Ala Ser Ser Ser Glu Lys Asn Glu Ser His Ser Ala Thr Lys Ser Ala Leu Ile Gln Lys Leu Met Gln Asp Ile Gln His Gln Asp Asn Tyr Glu Ala Ile Trp Glu Lys Ile Glu Gly Phe Val Asn Lys Leu Asp Glu Phe Ile Gln Trp Leu Asn Glu Ala Met Glu Thr Thr Glu Asn Trp Thr Pro Pro Lys Ala Glu Met Asp Asp Leu Lys Leu Tyr Leu Glu Thr His Leu Ser Phe Lys Leu Asn Val Asp Ser His Cys Ala WO 98J30695 PCT/US981~00543 Leu Lys Glu Ala Val Glu Glu Glu Gly His Gln Leu Leu Glu Leu Ile Ala Ser His Lys Ala Gly Leu Lys Asp Met Leu Arg Met Ile Ala Ser Gln Trp Lys Glu Leu Gln Arg Gln Ile Lys Arg Gln His Ser Trp Ile Leu Arg Ala Leu Asp Thr Ile Lys Ala Glu Ile Leu Ala Thr Asp Val Ser Val Glu Asp Glu Glu Gly Thr Gly Ser Pro Lys Ala Glu Val Gln Leu Cys Tyr Leu Glu Ala Gln Arg Asp Ala Val Glu Gln Met Ser Leu Lys Leu Tyr Ser Glu Gln Tyr Thr Ser Ser Ser Lys Arg Lys Glu Glu Phe Ala Asp Met Ser Lys Val His Ser Val Gly Ser Asn Gly Leu Leu Asp Phe Asp Ser Glu Tyr Gln Glu Leu Trp Asp Trp Leu Ile Asp Met Glu Ser Leu Val Met Asp Ser His Asp Leu Met Met Ser Glu Glu Gln Gln Gln His Leu Tyr Lys Val Arg Ala Thr Leu Pro Ala Phe Thr Leu Leu Trp Lys Ile (2) INFORMATION FOR SEQ ID N0:19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3442 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:19:

gg WO ~ PCT/US98/00543 AGATTTCTCC AAAAGTCCAG TTA,AAAAi9AA GTCTTTTGTT CAAGTTGGTG GGGATATTGT1440 TATTACCAGG TCAGATGCTG GATCATATAC ATGCATAGCC ACAAA'TCAGT TTGGCACTGC 1620 wo 9sr3o69s rcritrs9sroos43 AAAAAGGAGT

GGAATATTCA

GTTTATCTGC

AAGTGGAAGA

ACAGTCCCAT

CTGTTGCTAC

TGAGTCCTTG

AACCAAGTGA

TAAACATCCA

CAGAAGAACT

CGACAACCTG

ATGAAAGCAT

GAGAAGGATC

CCCCAAGGGG

CTATTGCCTG

ATGACTCCAA

ACATCACGGG

CTGGGACAGG

AACCACCAGC

ATGTAAAAAC

ACAGACAGAG

CATTTGAAGA

GCAGTGAGGA

TAGAACCTAG

CAGCCACTTA

TCTGTATATA

AAAGAAAAAA

AATTTTAAAC

ACTTAAAAAA Fl,~~i~AAAAAAA AA 3 4 4 2 (2) INFORMATION FOR SEQ ID N0:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1036 amino acids (B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYP&: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:20:
Met Arg Leu Leu Trp Lys Leu Val Ile Leu Leu Pro Leu Ile Asn Ser Ser Ala Gly Asp Gly Leu Leu Ser Arg Pro Ile Phe Thr Gln Glu Pro His Asp Val Ile Phe Pro Leu Asp Leu Ser Lys Ser Glu Val Ile Leu 35 40 q5 Asn Cys Ala Ala Asn Gly Tyr Pro Ser Pro His Tyr Arg Trp Lys Gln Asn Gly Thr Asp Ile Asp Phe Thr Met Ser Tyr His Tyr Arg Leu Asp Gly Gly Ser Leu Ala Ile Asn Ser Pro His Thr Asp Gln Asp Ile Gly Met Tyr Gln Cys Leu Ala Thr Asn Leu Leu Gly Thr Ile Leu Ser Arg Lys Ala Lys Leu Gln Phe Ala Tyr Ile Glu Asp Phe Glu Thr Lys Thr Arg Ser Thr Val Ser Val Arg Glu Gly Gln Gly Val Val Leu Leu Cys Gly Pro Pro Pro His Phe Gly Asp Leu Ser Tyr Ala Trp Thr Phe Asn Asp Asn Pro Leu Tyr Val Gln Glu Asp Asn Arg Arg Phe Val Ser Gln Glu Thr Gly Asn Leu Tyr Ile Ala Lys Val Glu Pro Ser Asp Val Gly Asn Tyr Thr Cys Phe Ile Thr Asn Lys Glu Ala Gln Arg Ser Val Gln Gly Pro Pro Thr Pro Leu Val Gln Arg Thr Asp Gly Val Met Gly Glu Tyr Glu Pro Lys Ile Glu Val Arg Phe Pro Glu Thr Ile Gln Ala Ala Lys Asp Ser Ser Val Lys Leu Glu Cys Phe Ala Leu Gly Asn Pro Val Pro Asp Ile Ser Trp Arg Arg Leu Asp Gly Ser Pro Leu Pro Gly Lys Val Lys Tyr Ser Lys Ser Gln Ala Ile Leu Glu Ile Pro Asn Phe Gln Gln Glu Asp Glu Gly Phe Tyr Glu Cys Ile Ala Ser Asn Leu Arg Gly Arg Asn Leu Ala Lys Gly Gln Leu Ile Phe Tyr Ala Pro Pro Glu Trp Glu Gln Lys Ile Gln Asn Thr His Leu Ser Ile Tyr Asp Asn Leu Leu Trp Glu Cys Lys Ala Ser Gly Lys Pro Asn Pro Trp Tyr Thr Trp Leu Lys Asn Gly Glu Arg Leu Asn Pro Glu Glu Arg Ile Gln Ile Glu Asn Gly Thr Leu Ile Ile Thr Met Leu Asn Val Ser Asp Ser Gly Val Tyr Gln Cys Ala Ala Glu Asn Lys Tyr Gln Ile Ile Tyr Ala Asn Ala Glu Leu Arg Val Leu Ala Ser Ala Pro Asp Phe Ser Lys Ser Pro Val Lys Lys Lys Ser Phe Val Gln Val Gly Gly Asp Ile Val Ile Gly Cys Lys Pro Asn Ala Phe Pro Arg Ala Ala Ile Ser Trp Lys Arg Gly Thr Glu Thr Leu Arg Gln Ser Lys Arg Ile Phe Leu Leu Glu Asp Gly Ser Leu Lys Ile Tyr Asn Ile Thr Arg Ser Asp Ala Gly Ser Tyr Thr Cys Ile Ala Thr Asn Gln Phe Gly Thr Ala Lys Asn Thr Gly Ser Leu Ile Val Lys Glu Arg Thr Val Ile Thr Val Pro Pro Ser Lys Met Asp Val Thr Val Gly Glu Ser Ile Val Leu Pro Cys Gln Val Ser His Asp Pro Ser Ile Glu Val Val Phe Val Trp Phe Phe Asn Gly Asp Val Ile Asp Leu Lys Lys Gly Val Ala His Phe Glu Arg Ile Gly Gly Glu Ser Val Gly ~ 545 550 555 560 Asp Leu Met Ile Arg Asn Ile Gln Leu His His Ser Gly Lys Tyr Leu Cys Thr Val Gln Thr Thr Leu GIu Ser Leu Ser Ala Val Ala Asp Ile Ile Val Arg Gly Pro Pro Gly Pro Pro Glu Asp Val Gln Val Glu Asp Ile Ser Ser Thr Thr Ser Gln Leu Ser Trp Arg Ala Gly Pro Asp Asn Asn Ser Pro Ile Gln Ile Phe Thr Ile Gln Thr Arg Thr Pro Phe Ser Val Gly Trp Gln Ala Val Ala Thr Val Pro Glu Ile Leu Asn Gly Lys Thr Tyr Asn Ala Thr Val Val Gly Leu Ser Pro Trp Val Glu Tyr Glu Phe Arg Val Val Ala Gly Asn Ser Ile Gly Ile Gly Glu Pro Ser Glu Pro Ser Glu Leu Leu Arg Thr Lys Ala Ser Val Pro Val Val Ala Pro Val Asn Ile His Gly Gly Gly Gly Ser Arg Ser Glu Leu Val Ile Thr Trp Glu Ser Ile Pro Glu Glu Leu Gln Asn Gly Glu Gly Phe Gly Tyr Ile Ile Met Phe Arg Pro Val Gly Ser Thr Thr Trp Ser Lys Glu Lys Val Ser Ser Val Glu Ser Ser Arg Phe Val Tyr Arg Asn Glu Ser Ile Ile Pro Leu Ser Pro Phe Glu Val Lys Val Gly Val Tyr Asn Asn Glu Gly Glu Gly Ser Leu Ser Thr Val Thr Ile Val Tyr Ser Gly Glu Asp Glu Pro Gln Leu Ala Pro Arg Gly Thr Ser Leu Gln Ser Phe Ser Ala Ser Glu Met Glu Val Ser Trp Asn Ala Ile Ala Trp Asn Arg Asn Thr Gly Arg Val Leu Gly Tyr Glu Val Leu Tyr Trp Thr Asp Asp Ser Lys Glu Ser Met Ile Gly Lys Ile Arg Val Ser Gly Asn Val Thr Thr Lys Asn Ile Thr Gly Leu Lys Ala Asn Thr Ile Tyr Phe Ala Ser Val Arg Ala Tyr Asn Thr Ala Gly Thr Gly Pro Ser Ser Pro Pro Val Asn Val Thr Thr Lys Lys Ser Pro Pro Ser Gln Pro Pro Ala Asn Ile Ala Trp Lys Leu Thr Asn Ser Lys Leu Cys Leu Asn Trp Glu His Val Lys Thr Met Glu Asn Glu Ser Glu Val Leu Gly Tyr Lys Ile Leu Tyr Arg Gln Asn Arg Gln Ser Lys Thr His Ile Leu Glu Thr Asn Asn Thr Ser Ala Glu Leu Leu Val Pro Phe Glu Glu Asp Tyr Leu Ile Glu Ile Arg Thr Val Ser Asp Gly Gly Asp Gly Ser Ser Ser Glu Glu Ile Arg Ile Pro Lys Met Ser Ser Leu Ser Ser Arg Gly Ile Gln Phe Leu Glu Pro Ser Thr His Phe Leu Ser Ile Val Ile Val Ile Phe Ser Leu Phe Cys Tyr Ser Ala Thr Tyr Leu Met Asn Lys Thr Ile Asn Leu (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) INFORMATION FOR SEQ ID N0:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID 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:

(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"

wo 9sr3o69s rcr~rs9s~oosa3 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:24:

(2) INFORMATION FOR SEQ ID N0:25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:25:

(2) INFORMATION FOR SEQ ID N0:26:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "oligonucleotide"
(xi) SEQUENCE DESCRIPTION: SEQ ID 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 (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 N0:28:

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

(2) INFORMATION FOR SEQ ID N0:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 29 base pairs (8) 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:
ANCAGTCTATC TTCCAGTCAG TATCTCAA 2g

Claims (37)

What is claimed is:

1. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:1 from nucleotide 79 to nucleotide 612;
(c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;
(d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
(e) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone BH272_3 deposited under accession number ATCC 98289;
(f) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:2;
(h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:2 having biological activity;
(i) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(f) above;
(j) a polynucleotide which encodes a species homologue of the protein of (g) or (h) above ; and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h).

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

3. A host cell transformed with a composition of claim 2.

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

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

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

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

8. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:2;
(b) the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164;
(c) fragments of the amino acid sequence of SEQ ID NO:2; and (d) the amino acid sequence encoded by the cDNA insert of clone BH272_3 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

10. The composition of claim 8, wherein said protein comprises the amino acid sequence of SEQ ID NO:2 from amino acid 13 to amino acid 164.

11. The composition of claim 8, further comprising a pharmaceutically acceptable carrier.

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

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

14. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 100 to nucleotide 675;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:5 from nucleotide 208 to nucleotide 489;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CH27_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:6;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:6 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above ; and (l) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

15. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:6;

(b) the amino acid sequence of SEQ ID NO:6 from amino acid 39 to amino acid 130;
(c) fragments of the amino acid sequence of SEQ ID NO:6; and (d) the amino acid sequence encoded by the cDNA insert of clone CH27_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

17. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 168 to nucleotide 1448;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:7 from nucleotide 440 to nucleotide 941;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CI542_2 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:8;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:8 having biological activity;
(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 capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

18. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:8;
(b) the amino acid sequence of SEQ ID NO:8 from amino acid 170 to amino acid 258;
(c) fragments of the amino acid sequence of SEQ ID NO:8; and (d) the amino acid sequence encoded by the cDNA insert of clone CI542_2 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

20. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 112 to nucleotide 1212;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 166 to nucleotide 1212;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:9 from nucleotide 704 to nucleotide 1033;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CN483_2 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;
(i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:10;
(j) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:10 having biological activity;
(k) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(h) above;
(l) a polynucleotide which encodes a species homologue of the protein of (i) or (j) above; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).

21. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:10;
(b) fragments of the amino acid sequence of SEQ ID NO:10; and (c) the amino acid sequence encoded by the cDNA insert of clone CN483_2 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

23. 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
NO:11 from nucleotide 234 to nucleotide 1466;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:11 from nucleotide 1 to nucleotide 367;

(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CO1224_2 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CO1224_2 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:12;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:12 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above; and (l) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

24. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:12;
(b) the amino acid sequence of SEQ ID NO:12 from amino acid 1 to amino acid 52;
(c) fragments of the amino acid sequence of SEQ ID NO:12; and (d) the amino acid sequence encoded by the cDNA insert of clone CO1224_2 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

26. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 845 to nucleotide 1102;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:13 from nucleotide 828 to nucleotide 1211;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CW768_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:14;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:14 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above; and (l) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

27. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:14;
(b) fragments of the amino acid sequence of SEQ ID NO:14; and (c) the amino acid sequence encoded by the cDNA insert of clone CW768_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

29. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 116 to nucleotide 1081;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:15 from nucleotide 509 to nucleotide 860;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CZ268_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:16;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:16 having biological activity;
(j) a polynucleotide which is an allelic variant of a polynucleotide of (a)-(g) above;
(k) a polynucleotide which encodes a species homologue of the protein of (h) or (i) above; and (l) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i).

30. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:16;
(b) the amino acid sequence of SEQ ID NO:16 from amino acid 153 to amino acid 248;
(c) fragments of the amino acid sequence of SEQ ID NO:16; and (d) the amino acid sequence encoded by the cDNA insert of clone CZ268_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

32. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 348 to nucleotide 2663;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:17 from nucleotide 397 to nucleotide 722;
(d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;
(e) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;
(f) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DH1308_1 deposited under accession number ATCC 98289;
(g) a polynucleotide encoding the mature protein encoded by the cDNA insert of done DH1308_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO:18;
(i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO:18 having biological activity;

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

33. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:18;
(b) the amino acid sequence of SEQ ID NO:18 from amino acid 19 to amino acid 125;
(c) fragments of the amino acid sequence of SEQ ID NO:18; and (d) the amino acid sequence encoded by the cDNA insert of clone DH1308_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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

35. A composition comprising an isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19;
(b) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 158 to nucleotide 3268;
(c) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 215 to nucleotide 3268;
(d) a polynucleotide comprising the nucleotide sequence of SEQ ID
NO:19 from nucleotide 55 to nucleotide 379;
(e) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;
(f) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;

(g) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone DL185_1 deposited under accession number ATCC 98289;
(h) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;
(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;
(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 capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j).

36. A composition comprising a protein, wherein said protein comprises an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:20;
(b) the amino acid sequence of SEQ ID NO:20 from amino acid 1 to amino acid 74;
(c) fragments of the amino acid sequence of SEQ ID NO:20; and (d) the amino acid sequence encoded by the cDNA insert of clone DL185_1 deposited under accession number ATCC 98289;
the protein being substantially free from other mammalian proteins.

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