JP2002504306A - Secreted proteins and polynucleotides encoding them - Google Patents

Abstract

  (57) [Summary] Disclosed are novel polynucleotides and the proteins encoded by them.

Description

DETAILED DESCRIPTION OF THE INVENTION

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

BACKGROUND proteinaceous factors invention (e.g., lymphokines, interferons, including cytokine such as CSFs and interleukins) has methods for the purpose of finding this 10
Matured rapidly over the years. Currently, conventional hybridization and expression cloning methods rely on information directly related to the discovered protein (ie,
In the case of hybridization cloning, depending on the partial DNA / amino acid sequence of the protein; in the case of expression cloning, the activity of the protein).
The new polypeptide is cloned "directly". Signal sequence cloning (
DNA sequences are isolated based on the presence of the currently well-recognized secretory leader sequence motifs), as well as more recent "indirect" cloning methods, such as various PCR or low stringency hybridization cloning methods. A large number of DNA / amino acid sequences for proteins known to have biological activity due to their secreted nature in the case of cloning of the leader sequence, or due to the cell or tissue source in the case of the PCR method. Has made progress by enabling. The present invention is directed to these proteins and the polynucleotides that encode them.

[0003] In one embodiment of the invention, the invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) comprising the nucleotide sequence of SEQ ID NO: 1 (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 from nucleotide 54 to nucleotide 1283; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 from nucleotide 648 to nucleotide 1283; (d) A polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 from nucleotide 458 to nucleotide 947; (e) clone AY4212, deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of (a) clone AY4212 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of (c) clone AY4212 deposited under accession number ATCC 98145.
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of (h) clone AY4212, deposited under accession number ATCC 98145.
A polynucleotide encoding a mature protein encoded by the cDNA insert of (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 2; (j) a biologically active SEQ ID NO: A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of (2); (k) a polynucleotide which is an allelic variant of the polynucleotide of (a) to (h); A) a polynucleotide that encodes a species homolog of the protein; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such a polynucleotide comprises a nucleotide sequence from nucleotide 54 to nucleotide 1283 of SEQ ID NO: 1; a nucleotide sequence from nucleotide 648 to nucleotide 1283 of SEQ ID NO: 1; Nucleotide sequence; accession number AT
The nucleotide sequence of the full length protein coding sequence of clone AY4212, deposited as CC98145; or the mature protein coding sequence of clone AY4212, deposited as accession number ATCC98145. In another preferred embodiment, the polynucleotide has accession number ATCC 98.
145 encodes the full-length or mature protein encoded by the cDNA insert of clone AY4212 deposited as 145. In yet another preferred embodiment, the invention provides a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 180 to amino acid 298 of SEQ ID NO: 2. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 1. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (b) an amino acid sequence from amino acid 180 to amino acid 298 of SEQ ID NO: 2; (c) a fragment of the amino acid sequence of SEQ ID NO: 2; and (d) a deposit of accession number ATCC 98145. Clone AY421 2 cloned
Amino acid sequence encoded by the cDNA insert of FIG. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 2 or the amino acid sequence from amino acid 180 to amino acid 298 of SEQ ID NO: 2.

[0004] In one embodiment, the invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 3; (B) a polynucleotide comprising SEQ ID NO: 3 from nucleotide 720 to nucleotide 974; (c) a polynucleotide comprising SEQ ID NO: 3 from nucleotide 715 to nucleotide 947; (d) clone BV278 deposited under accession number ATCC 98145 2
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of (c) BV2782 deposited under accession number ATCC 98145
A polynucleotide encoding the full length protein encoded by the cDNA insert of (b) clone BV2782 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of: (g) clone BV2782 deposited under accession number ATCC 98145.
(H) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 4; (i) an amino acid sequence of SEQ ID NO: 4 having biological activity (J) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; (k) a species homology 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). Preferably, such a polynucleotide is nucleotide 720 of SEQ ID NO: 3.
Nucleotide sequence from nucleotide 715 to nucleotide 947 of SEQ ID NO: 3; accession number ATCC98
145, containing the nucleotide sequence of the full length protein coding sequence of clone BV2782 deposited as 145; or the mature protein coding sequence of clone BV2782 deposited as accession number ATCC 98145. In another preferred embodiment, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone BV2782, deposited under accession number ATCC 98145. In yet another preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of amino acid 1 to amino acid 76 of SEQ ID NO: 4. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 3. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (b) an amino acid sequence from amino acid 1 to amino acid 76 of SEQ ID NO: 4; (c) a fragment of the amino acid sequence of SEQ ID NO: 4; and (d) a deposit under accession number ATCC 98145. Clone BV278 2
Amino acid sequence encoded by the cDNA insert of FIG. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 4 or the amino acid sequence from amino acid 1 to amino acid 76 of SEQ ID NO: 4.

[0005] In one embodiment, the invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 5; (B) a polynucleotide comprising the nucleotide sequence from nucleotide 36 to nucleotide 968 of SEQ ID NO: 5; (c) a polynucleotide comprising the nucleotide sequence from nucleotide 340 to nucleotide 717 of SEQ ID NO: 5; (d) accession number ATCC 98145 A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone C5441 deposited as accession number; (e) a polynucleic acid encoding the full-length protein encoded by the cDNA insert of clone C5441 deposited under accession number ATCC 98145 (F) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone C5441 deposited under accession number ATCC98145; (g) the mature protein encoded by the cDNA insert of clone C5441 deposited under accession number ATCC98145 (H) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 6; (i) a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 6 having biological activity (J) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; and (k) encodes a species homolog of the protein of (h) or (i) above. A polynucleotide that is; and (l) under stringent conditions A polynucleotide capable of hybridizing to any one of the polynucleotides shown in (a) to (i). Preferably, such a polynucleotide is a nucleotide sequence from nucleotide 36 to nucleotide 968 of SEQ ID NO: 5; nucleotide 3 of SEQ ID NO: 5
Nucleotide sequence from 40 to nucleotide 717; accession number ATCC 981
Nucleotide sequence of the full length protein coding sequence of clone C544_1 deposited as accession number 45; or clone C deposited as accession number ATCC 98145
5441 contains the nucleotide sequence of the mature protein coding sequence. In another preferred embodiment, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone C5441 deposited under accession number ATCC 98145. In still other preferred embodiments, the present invention provides
Provided is a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 103 to amino acid 227 of SEQ ID NO: 6. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 5. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (b) an amino acid sequence from amino acid 103 to amino acid 227 of SEQ ID NO: 6; (c) a fragment of the amino acid sequence of SEQ ID NO: 6; and (d) a deposit of Accession No. ATCC 98145. Amino acid sequence encoded by the cloned cDNA insert of clone C5441. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 6 or the amino acid sequence from amino acid 103 to amino acid 227 of SEQ ID NO: 6.

[0006] In one embodiment, the 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: 7; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 7 from nucleotide 151 to nucleotide 1398; (c) a polynucleotide comprising the nucleotide sequence of nucleotide 637 to nucleotide 1398 of SEQ ID NO: 7; (d) SEQ ID NO: A polynucleotide comprising a nucleotide sequence from nucleotide 255 to nucleotide 429 of 7; (e) clone CC332 deposited under accession number ATCC 98145;
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of SEQ ID NO: 3; (f) clone CC332 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of No. 3; (g) clone CC332 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of 3; (h) clone CC332 3 deposited under accession number ATCC 98145
A polynucleotide encoding a mature protein encoded by the cDNA insert of SEQ ID NO: 3; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 8; (j) a SEQ ID NO having biological activity : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of 8; (k) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (h) above; (l) a (i) or (i) j) a polynucleotide encoding a species homolog of the protein; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(j). Preferably, such a polynucleotide is nucleotide 151 of SEQ ID NO: 7.
Nucleotide sequence from nucleotide 637 to nucleotide 1398 of SEQ ID NO: 7; nucleotide sequence from nucleotide 255 to nucleotide 429 of SEQ ID NO: 7; Accession No. A
The nucleotide sequence of the full-length protein coding sequence of clone CC33233 deposited as TCC98145; or the mature protein coding sequence of clone CC33233 deposited as accession number ATCC98145. In another preferred embodiment, the polynucleotide has accession number ATC.
Encodes the full-length or mature protein encoded by the cDNA insert of clone CC33233 deposited as C98145. In yet another preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence of amino acid 36 to amino acid 93 of SEQ ID NO: 8. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 7. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: a) the amino acid sequence of SEQ ID NO: 8; (b) the amino acid sequence from amino acid 36 to amino acid 93 of SEQ ID NO: 8; (c) a fragment of the amino acid sequence of SEQ ID NO: 8; and (d) the accession number ATCC 98145. Clone CC332 3 cloned
Amino acid sequence encoded by the cDNA insert of No. 3. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 8 or the amino acid sequence from amino acid 36 to amino acid 93 of SEQ ID NO: 8.

In one embodiment, the 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: 9; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 9 from nucleotide 89 to nucleotide 436; (c) a polynucleotide comprising the nucleotide sequence of nucleotide 212 to nucleotide 436 of SEQ ID NO: 9; (d) accession number ATCC 98145 Clone CC365 4 deposited as
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of SEQ ID NO: 0; (e) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide encoding the full length protein encoded by the cDNA insert of No. 0; (f) clone CC3654 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of SEQ ID NO: 0; (g) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide encoding a mature protein encoded by a cDNA insert of 0; (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 10; (i) a SEQ ID NO having biological activity (J) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g); (k) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; a polynucleotide encoding a species homolog of the protein of i); and (l) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i). Preferably, such a polynucleotide is a nucleotide sequence from nucleotide 89 to nucleotide 436 of SEQ ID NO: 9; nucleotide 2 of SEQ ID NO: 9
Nucleotide sequence from 12 to nucleotide 436; accession number ATCC 981
45 contains the nucleotide sequence of the full length protein coding sequence of clone CC36540 deposited as No. 45; or the mature protein coding sequence of clone CC36540 deposited as Accession No. ATCC 98145. In another preferred embodiment, the polynucleotide has accession number ATCC 98145.
And encodes the full-length or mature protein encoded by the cDNA insert of clone CC36540 deposited as In yet another preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 10. Other embodiments provide genes corresponding to the cDNA sequences of SEQ ID NO: 9 and SEQ ID NO: 11. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins. A) an amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 10; (c) a fragment of the amino acid sequence of SEQ ID NO: 10; and (d) an accession number ATCC 98145. Clone CC365 4
Amino acid sequence encoded by 0 cDNA insert. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 10 or the amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 10.

[0008] 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: 32; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 32 from nucleotide 21 to nucleotide 410; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 32 from nucleotide 144 to nucleotide 410; (d) SEQ ID NO: A polynucleotide comprising a nucleotide sequence of from nucleotide 68 to nucleotide 368 of 32; (e) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of SEQ ID NO: 0; (f) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide encoding the full-length protein encoded by the cDNA insert of No. 0; (g) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of SEQ ID NO: 0; (h) clone CC365_4 deposited under accession number ATCC 98145
A polynucleotide encoding a mature protein encoded by a cDNA insert of 0; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 33; (j) a SEQ ID NO having biological activity : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33
(K) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (h) above; (l) a species homology 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 a polynucleotide is nucleotide 21 of SEQ ID NO: 32.
Nucleotide sequence from nucleotide 144 to nucleotide 410 of SEQ ID NO: 32; nucleotide sequence from nucleotide 68 to nucleotide 368 of SEQ ID NO: 32; accession number AT
The nucleotide sequence of the full length protein coding sequence of clone CC36540 deposited as CC98145; or the mature protein coding sequence of clone CC36540 deposited as accession number ATCC98145. In another preferred embodiment, the polynucleotide has accession number ATCC.
It encodes the full-length or mature protein encoded by the cDNA insert of clone CC36540 deposited as 98145. In a further preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 33. In a further preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33 having biological activity, preferably the fragment comprises 8 (more preferably 20) , Most preferably 30)
The present invention also provides a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33 having biological activity, said fragment comprising the contiguous amino acids of SEQ ID NO: 33. It contains the amino acid sequence from amino acid 60 to amino acid 69. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 32. A further embodiment of the invention provides a method for producing an isolated polynucleotide, wherein the method is selected from the group consisting of: (a) a method comprising the steps of: (i) 65 in 6X SSC. Preparing one or more polynucleotide probes that hybridize at 0 ° C. to a nucleotide sequence selected from the group consisting of: (aa) SEQ ID NO: 32 (at the 3 ′ end of SEQ ID NO: 32) (Except poly (A) tail); and (ab) clone CC36540 deposited as ATCC 98145
(Ii) hybridizing the probe (s) to human DNA; (iii) isolating the DNA polynucleotide detected with the probe (s); and (B) a method comprising the steps of: (i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 ° C. to a nucleotide sequence selected from the group consisting of: (Ba) SEQ ID NO: 32 (excluding the poly (A) tail at the 3 ′ end of SEQ ID NO: 32); and (bb) Clone CC36540 deposited as ATCC 98145
(Ii) hybridizing the primer (s) to human DNA; (iii) amplifying the human DNA sequence; then (iv) the polynucleotide of step (b) (iii) Isolating the product. Preferably, the nucleotide sequence of the polynucleotide isolated by this method corresponds to the cDNA sequence of SEQ ID NO: 32, and the 5 ′ of SEQ ID NO: 32
It is a continuous sequence from the nucleotide sequence corresponding to the terminus to the nucleotide sequence corresponding to the 3 ′ terminus of SEQ ID NO: 32, except that the poly (
A) The tail is removed. Also, preferably, the polynucleotide isolated by the above method comprises a nucleotide sequence corresponding to nucleotide 21 to nucleotide 410 of the cDNA sequence of SEQ ID NO: 32, and comprises the nucleotide sequence from nucleotide 21 to nucleotide 410 of SEQ ID NO: 32. Contiguous from the nucleotide sequence corresponding to the 5 'end of the sequence to the nucleotide sequence corresponding to the 3' end of the sequence from nucleotide 21 to nucleotide 410 of SEQ ID NO: 32. Also preferably, the polynucleotide isolated by the above step comprises a nucleotide sequence corresponding to the cDNA sequence from nucleotide 144 to nucleotide 410 of SEQ ID NO: 32, and from nucleotide 144 to nucleotide 410 of SEQ ID NO: 32
From the nucleotide sequence corresponding to the 5 'end of the sequence to the nucleotide sequence corresponding to the 3' end of the sequence from nucleotide 144 to nucleotide 410 of SEQ ID NO: 32. Also preferably, the polynucleotide isolated by the above method comprises a nucleotide sequence corresponding to the cDNA sequence from nucleotide 68 to nucleotide 368 of SEQ ID NO: 32;
A sequence from the nucleotide sequence corresponding to the 5 'end of the sequence from nucleotide 68 to nucleotide 368 of 32 to the nucleotide sequence corresponding to the 3' end of the sequence from nucleotide 68 to nucleotide 368 of SEQ ID NO: 32 It is. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: a) the amino acid sequence of SEQ ID NO: 33; (b) the amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 33; (c) the amino acid sequence of SEQ ID NO: 33, comprising eight consecutive amino acids of SEQ ID NO: 33 A fragment of the amino acid sequence; and (d) clone CC365_4 deposited under accession number ATCC 98145.
Amino acid sequence encoded by 0 cDNA insert. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 33. In a further preferred embodiment,
The present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33 having biological activity, preferably the fragment comprises eight consecutive amino acids of SEQ ID NO: 33. Alternatively, the present invention provides a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33 having biological activity, preferably, the fragment comprises the amino acid sequence of amino acids 60 to 69 of SEQ ID NO: 33.

[0009] In one embodiment, the invention provides a composition comprising an isolated polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 12; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 12 from nucleotide 769 to nucleotide 966; (c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone CG684 deposited under accession number ATCC 98145; A) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CG684 deposited under accession number ATCC 98145; (F) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone CG684 deposited under accession number ATCC 98145; (g) comprising the amino acid sequence of SEQ ID NO: 13 A polynucleotide encoding a protein; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 13 having biological activity; (i) a polynucleotide encoding the above-mentioned (a) to (f) (J) a polynucleotide that encodes a species homolog of the protein of (g) or (h) above; and (k) under stringent conditions (a)-( h) a polynucleotide capable of hybridizing to any one of the polynucleotides set forth in h) Preferably, such a polynucleotide is nucleotide 76 of SEQ ID NO: 12.
Nucleotide sequence from 9 to nucleotide 966; accession number ATCC 9814
Nucleotide sequence of the full length protein coding sequence of clone CG684 deposited as accession number 5; or clone CG deposited as accession number ATCC 98145.
684 contains the nucleotide sequence of the mature protein coding sequence. In another preferred embodiment, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone CG684 deposited under accession number ATCC 98145. In yet another preferred embodiment, the invention provides a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 18 to amino acid 57 of SEQ ID NO: 13. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 12. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (b) amino acid sequence from amino acid 18 to amino acid 57 of SEQ ID NO: 13; (c) a fragment of the amino acid sequence of SEQ ID NO: 13; and (d) deposited as accession number ATCC 98145. Amino acid sequence encoded by the cloned cDNA insert of clone CG684. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 13 or the amino acid sequence from amino acid 18 to amino acid 57 of SEQ ID NO: 13.

[0010] In one embodiment, the 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: 14; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 14 from nucleotide 1042 to nucleotide 1389; (c) a polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of clone D3291 deposited under accession number ATCC 98145; A) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145; (F) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145; (g) a polynucleotide comprising the amino acid sequence of SEQ ID NO: 15 A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 15 having biological activity; (i) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 15 having biological activity; (J) a polynucleotide encoding a species homolog of the protein of (g) or (h) above; and (k) under stringent conditions Polynucleotide capable of hybridizing to any one of the polynucleotides shown in (h) De. Preferably, such a polynucleotide is nucleotide 10 of SEQ ID NO: 14.
Nucleotide sequence from 42 to nucleotide 1389; accession number ATCC 98
145, including the nucleotide sequence of the full length protein coding sequence of clone D3291 deposited as 145; or the mature protein coding sequence of clone D3291 deposited as accession number ATCC 98145. In another preferred embodiment, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 14. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (b) a fragment of the amino acid sequence of SEQ ID NO: 15; and (c) the amino acid sequence encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 15.

[0011] 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: 18; (B) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 18 from nucleotide 279 to nucleotide 515; (c) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone H6983 deposited under accession number ATCC 98145; A) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145; (e) a mature protein coding sequence for clone H6983 deposited under accession number ATCC 98145 (F) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145; (g) comprising the amino acid sequence of SEQ ID NO: 19 A polynucleotide encoding a protein; (h) a polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 19 having biological activity; (i) a polynucleotide encoding a protein (J) a polynucleotide that encodes a species homolog of the protein of (g) or (h) above; and (k) under stringent conditions (a)-( h) a polynucleotide capable of hybridizing to any one of the polynucleotides set forth in h) Preferably, such a polynucleotide is nucleotide 27 of SEQ ID NO: 18.
Nucleotide sequence from 9 to nucleotide 515; accession number ATCC 9814
Nucleotide sequence of the full length protein coding sequence of clone H6983 deposited as accession number 5; or clone H6 deposited as accession number ATCC 98145.
983 contains the nucleotide sequence of the mature protein coding sequence. In another preferred embodiment, the polynucleotide encodes the full-length or mature protein encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145. Other embodiments provide genes corresponding to the cDNA sequences of SEQ ID NO: 17 and SEQ ID NO: 18. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: a) the amino acid sequence of SEQ ID NO: 19; (b) a fragment of the amino acid sequence of SEQ ID NO: 19; and (c) the amino acid sequence encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 19.

In one embodiment, the 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: 21; (B) a polynucleotide comprising the nucleotide sequence from nucleotide 199 to nucleotide 1155 of SEQ ID NO: 21; (c) a polynucleotide comprising the nucleotide sequence from nucleotide 304 to nucleotide 1155 of SEQ ID NO: 21; (d) accession number ATCC 98145 Clone H9620 deposited as
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of (c) H96320 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of (h) clone H96320 deposited under accession number ATCC 98145;
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of (c) H96320 deposited under accession number ATCC 98145.
A polynucleotide encoding a mature protein encoded by the cDNA insert of (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 22; (i) a SEQ ID NO: having biological activity: A polynucleotide encoding a protein comprising a fragment of 22 amino acid sequences; (j) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; (k) a (h) or (i) A) a polynucleotide that encodes a species homolog of the protein of claim 1); and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a) to (i). Preferably, such a polynucleotide is nucleotide 19 of SEQ ID NO: 21.
Nucleotide sequence from 9 to nucleotide 1155; nucleotide sequence from nucleotide 304 to nucleotide 1155 of SEQ ID NO: 21; accession number AT
The full length protein coding sequence of clone H96320 deposited as CC98145; or the mature protein coding sequence of clone H96320 deposited as accession number ATCC 98145. In another preferred embodiment, the polynucleotide has accession number ATCC 98.
145 encodes the full-length or mature protein encoded by the cDNA insert of clone H96320 deposited as 145. In yet another preferred embodiment, the present invention provides a polynucleotide encoding a protein comprising the amino acid sequence from amino acid 19 to amino acid 84 of SEQ ID NO: 22. Another embodiment provides a gene corresponding to the cDNA sequence of SEQ ID NO: 21. In another embodiment, the present invention provides a composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: a) the amino acid sequence of SEQ ID NO: 22; (b) the amino acid sequence from amino acid 19 to amino acid 84 of SEQ ID NO: 22; (c) a fragment of the amino acid sequence of SEQ ID NO: 22; and (d) the accession number ATCC 98145. Clone H9620
Amino acid sequence encoded by the cDNA insert of FIG. Preferably, such a protein comprises the amino acid sequence of SEQ ID NO: 22 or the amino acid sequence from amino acid 19 to amino acid 84 of the amino acid sequence of SEQ ID NO: 22.

[0013] In certain preferred embodiments, the polynucleotide is operably linked to an expression control sequence. The invention also provides host cells, including bacterial, yeast, insect and mammalian cells, transformed with such polynucleotide compositions. Also,
An organism having enhanced, reduced, or modified expression of a gene corresponding to a polynucleotide sequence disclosed herein is provided by the invention. Further, a method for producing a protein, comprising: (a) growing a culture of a host cell transformed with such a polynucleotide composition in a suitable medium; and (b) purifying the protein from the culture. There is also provided a method comprising: The protein produced by such a method is also provided by the present invention. Preferred specific examples include those in which the protein produced by such a method is a mature form of the protein. The protein composition of the present invention may further contain a pharmaceutically acceptable carrier. Compositions comprising antibodies specifically reactive with such proteins are also provided by the present invention. Also provided is a method for preventing, treating or ameliorating a medical condition comprising administering to a mammalian subject a therapeutically effective amount of a composition comprising a protein of the invention and a pharmaceutically acceptable carrier.

DETAILED DESCRIPTION Isolated Proteins and Polynucleotides The nucleotide and amino acid sequences for each of the clones and proteins disclosed herein are reported below. By sequencing the deposited clones by known methods, the actual nucleotide sequence of each clone can be readily determined. The deduced amino acid sequence (both full length as well as mature) can then be determined from such nucleotide sequences. By expressing the clones in a suitable host cell, collecting the proteins, and then determining the sequence, the amino acid sequence of the protein encoded by each clone can also be determined. For each protein disclosed, Applicants have identified what has been determined to be the best identifiable reading frame using sequence information available at the time of filing. As used herein, the term "secreted" protein refers to that which is transported across a membrane when expressed in a suitable host cell, including transport as a result of a signal sequence in its amino acid sequence. “Secreted” proteins include, but are not limited to, proteins that are totally secreted (eg, soluble proteins) or partially secreted (eg, receptors) from the cell in which they are expressed. "Secreted" proteins also include, but are not limited to, those that are transported across the endoplasmic reticulum membrane. Clone "AY421 2" The polynucleotide of the present invention has been identified as clone AY421-2. A
Y4212 is isolated from an adult retinal cDNA library using methods selective for the cDNA encoding the secreted protein (see US Pat. No. 5,536,637), or for computer analysis of the amino acid sequence of the encoded protein. On the basis of,
It was identified as encoding a secreted or transmembrane protein. AY4212 is a full-length clone and is a secreted protein (also referred to herein as "AY4212 protein").
Contains the entire coding sequence. The nucleotide sequence of AY4212 determined this time is shown in SEQ ID NO: 1. This time, the applicant considers the correct reading frame and deduced amino acid sequence of AY4212 protein corresponding to the above nucleotide sequence to be shown in SEQ ID NO: 2. Amino acids 186 to 198 are a predicted leader / signal sequence, the predicted mature amino acid sequence starts at amino acid 199, or amino acids 186 to 198 are a transmembrane domain. EcoRI / No obtainable from the deposit containing clone AY4212
The tI restriction fragment should be approximately 1500 bp in length. The deduced amino acid sequence disclosed herein for AY4212 was compared to BLASTN / B
Searches were made against GenBank and GenSeq nucleotide sequence databases using the LASTX and FASTA search protocols. AY4212 is compatible with U46493 (Cloning vect
or pFlp recombinase gene, complete cds) showed at least some similarity to the sequence identified. The deduced amino acid sequence disclosed herein for AY4212 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative AY421 2 protein is AA016992 (ze
33d02.r1 Soares retina N2b4HR Homo sapiens cDNA clone 360771 5 'similar
to WP T06D8.5 CE02326 YER141W), H60299 (yr41b09.r1 Homo sapiens cDNA clo
ne 207833 5 '), L38643 (Saccharomyces cerevisiae cytochrome oxidase assem
bly), N25978 (yx88b04.s1 Homo sapiens cDNA clone 268783 3 '), and R598
It showed at least some similarity to the sequence identified as 51 (yh07a09.r1 Homo sapiens cDNA). The deduced amino acid sequence disclosed herein for AY4212 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative AY421 2 protein is L38643 (YSCCOX15A_
1 cytochrome oxidase assembly factor [Saccharomyces cerevisiae]) and
It showed at least some similarity to the sequence identified as Z49130 (CET06D8_5 T06D8.5 [Caenorhabditis elegans]). Based on sequence similarity, AY4
The 212 protein and each similar protein or peptide may share at least some activity. According to the TopPredII computer program, in the AY4221 protein sequence, amino acids 80, 193, 237, 274 of SEQ ID NO: 2
Six potential transmembrane domains centered around 336 and 376 are expected.

Clone "BV2782" The polynucleotide of the present invention has been identified as clone BV2782. B
V2782 was isolated from an adult brain cDNA library using methods selective for cDNAs encoding secreted proteins (see US Pat. No. 5,536,637).
Alternatively, it was identified as encoding a secreted protein or a transmembrane protein based on computer analysis of the amino acid sequence of the encoded protein. BV2782 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "BV2782 protein"). The nucleotide sequence of BV2782 determined this time is shown in SEQ ID NO: 3. This time, the applicant considers the correct reading frame and deduced amino acid sequence of BV2782 protein corresponding to the above nucleotide sequence to be shown in SEQ ID NO: 4. EcoRI / No obtainable from the deposit containing clone BV2782
The tI restriction fragment should be approximately 2300 bp in length. The nucleotide sequence of BV2782 disclosed herein was searched against the GenBank and GeneSeq databases using BLASTA / BLASTX and FASTA search protocols. BV278 2 is AA359704 (EST68845 Fetal lung II Homo sa
piens cDNA 5 'end) showed at least some similarity to the sequence identified. Based on sequence similarity, the BV2782 protein and each similar protein or peptide may share at least some activity. According to the TopPredII computer program, a potential transmembrane domain centering around amino acid 19 of SEQ ID NO: 4 in the BV2782 protein sequence is expected.

Clone "C544-1" The polynucleotide of the present invention has been identified as clone C544-1. C5
No. 441 discloses a method selective for cDNA encoding a secretory protein (US Pat.
No. 5,366,637), isolated from adult blood (phytohemagglutinin and phorbol myristate acetate and peripheral blood mononuclear cells treated with mixed lymphocyte reaction) cDNA library, or the amino acid sequence of the encoded protein. Based on computer analysis, it was identified as encoding a secreted or transmembrane protein. C544-1 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "C544-1 protein"). The nucleotide sequence of C5441 determined this time is shown in SEQ ID NO: 5. SEQ ID NO: 6 shows what the applicant currently considers to be the correct reading frame and deduced amino acid sequence for C5441 protein corresponding to the above nucleotide sequence. EcoRI / Not obtainable from the deposit containing clone C5441
The I restriction fragment should be approximately 1250 bp in length. The nucleotide sequence of C5441 disclosed herein was searched against the GenBank and GeneSeq databases using BLASTA / BLASTX and FASTA search protocols. C544 1 is AA029713 (ze95f03.s1 Soares fetal heart NbHH
19W Homo sapiens cDNA clone 366749 3 '), T53653 (ya98b10.s1 Homo sapiens
cDNA clone 69691 3 '), T85425 (yd76d07.r1 Homo sapiens cDNA clone 114157
5 '), and Z97634 (Human DNA sequence *** SEQUENCING IN PROGRESS *** fr
om clone 367G8; showed at least some similarity to the sequence identified as HTGS phase 1). Based on sequence similarity, the C5441 protein and each similar protein or peptide may share at least some activity. To
According to the pPredII computer program, amino acids 80, 110, 150, 170, 190, 220 and 260 of SEQ ID NO: 6 in the C5441 protein sequence
Seven potential transmembrane domains centered around are expected. Also C544
One protein contains an EGF domain near amino acid 50 of SEQ ID NO: 6. A single EGF domain is present in human teratocarcinoma-derived growth factor 1, tissue plasminogen activator (TPA) and other proteins including but not limited to clotting factors VII, IX and X It is shown. A common feature of EGF repeats is that they are found in the extracellular domain of membrane bound proteins or proteins known to be secreted.

Clone "CC332 33" The polynucleotide of the present invention has been identified as clone CC33233.
CC332 33 can be isolated from an adult brain cDNA library using methods selective for the cDNA encoding the secreted protein (see US Pat. No. 5,536,637), or used for computer analysis of the amino acid sequence of the encoded protein. Based on this, it was identified as encoding a secreted or transmembrane protein. CC332 33
Is a full-length clone, containing the entire coding sequence of a secreted protein (also referred to herein as "CC33233 protein"). The nucleotide sequence of CC33233 determined this time is shown in SEQ ID NO: 7.
This time, the applicant considers the correct reading frame and deduced amino acid sequence of CC33233 protein corresponding to the above nucleotide sequence to be shown in SEQ ID NO: 8. Amino acid 1
50 to 162 is the predicted leader / signal sequence, the predicted mature amino acid sequence starts at amino acid 163, or amino acids 150 to 162 is the transmembrane domain. EcoRI / N obtainable from the deposit containing clone CC33233
The otI restriction fragment should be approximately 4400 bp in length. The nucleotide sequence of CC33233 disclosed herein was searched against the GenBank and GeneSeq databases using the BLASTA / BLASTX and FASTA search protocols. CC332 33 was obtained from N63467 (yy61d06.s1 Homo sapiens cDNA).
It showed at least some similarity to the sequence identified as clone 278027 3 '). According to the TopPredII computer program, a potential transmembrane domain centering around amino acid 10 of SEQ ID NO: 8 in the CC332 33 protein sequence is expected.

Clone "CC36540" The polynucleotide of the present invention has been identified as clone "CC36540" .
CC36540 is isolated from an adult brain cDNA library using methods selective for the cDNA encoding the secreted protein (see US Pat. No. 5,536,637), or for computer analysis of the amino acid sequence of the encoded protein. Based on this, it was identified as encoding a secreted or transmembrane protein. CC365 40
Is a full-length clone, containing the entire coding sequence of a secreted protein (also referred to herein as "CC36540 protein"). The nucleotide sequence of the 5 'portion of CC36540 determined this time is represented by SEQ ID NO:
It is shown in FIG. The reading frame for the coding region of SEQ ID NO: 9 is shown in SEQ ID NO: 10. The deduced amino acid sequence of CC36540 protein corresponding to the above nucleotide sequence is shown in SEQ ID NO: 10. Amino acids 29 to 41 of SEQ ID NO: 10 are a predicted leader / signal sequence, with the predicted mature amino acid sequence beginning at amino acid 42. Due to the hydrophobic nature of the putative leader / signal sequence, the putative leader / signal may act as a transmembrane domain without separating from the rest of the CC36540 protein. An additional nucleotide sequence from the 3 'portion of CC36540, including the poly A tail, is shown in SEQ ID NO: 11. The nucleotide sequence of CC36540 determined this time is shown in SEQ ID NO: 32, which contains a poly (A) tail. CC corresponding to the above nucleotide sequence
SEQ ID NO: 33 shows what the applicant currently considers to be the correct reading frame and deduced amino acid sequence for the 36540 protein. Amino acids 29 to 41 of SEQ ID NO: 33 are a predicted leader / signal sequence;
Start with. Due to the hydrophobic nature of the putative leader / signal sequence, the putative leader / signal may act as a transmembrane domain without separating from the rest of the CC36540 protein. EcoRI / N obtainable from the deposit containing clone CC36540
The otI restriction fragment should be approximately 1380 bp in length. The nucleotide sequence of CC36540 disclosed herein was searched against the GenBank and GeneSeq databases using the BLASTA / BLASTX and FASTA search protocols. CC36540 is H33410 (EST109372 Rattus sp. CDNA 5 '
end) and W21840 (58b3 Human retina cDNA Tsp509I-cleaved sublibrary H
omo sapiens cDNA) showed at least some similarity to the sequence identified. The deduced amino acid sequence disclosed herein for CC36540 was compared to BLASTX
A search was performed against the GenPept and GeneSeq amino acid sequence databases using a search protocol. The putative CC36540 protein is U13625 (cytochrome b [Pezoporu
s wallicus]) showed at least some similarity to the sequence identified as Based on sequence similarity, the CC36540 protein and each similar protein or peptide may share at least some activity.

Clone "CG684" The polynucleotide of the present invention has been identified as clone "CG684" . CG
684 describes a method selective for cDNAs encoding secreted proteins (US Pat.
No. 5,366,637), and was identified as encoding a secreted or transmembrane protein based on computer analysis of the amino acid sequence of the encoded protein. CG684 is a full-length clone and contains the entire coding sequence of a secreted protein (also referred to herein as "CG684 protein"). The nucleotide sequence of CG684 determined this time is shown in SEQ ID NO: 12. This time, the applicant considers the correct reading frame and deduced amino acid sequence of CG684 protein corresponding to the above nucleotide sequence to be shown in SEQ ID NO: 13. EcoRI / Not obtainable from the deposit containing clone CG684
The I restriction fragment should be approximately 1080 bp long. The nucleotide sequence of CG684 disclosed herein was searched against the GenBank and GeneSeq databases using BLASTA / BLASTX and FASTA search protocols. CG684 is M29854 (interleukin 4 receptor (IL-4) precur
It showed at least some similarity to the sequence identified as sor). Based on sequence similarity, the CG684 protein and each similar protein or peptide may share at least some activity.

Clone "D329-1" The polynucleotide of the present invention has been identified as clone D329-1. D3
291 describes a method selective for cDNA encoding a secretory protein (US Pat.
No. 5,366,637) isolated from adult blood (peripheral blood mononuclear cells treated with concanavalin A and phorbol myristate acetate) cDNA library or based on computer analysis of the amino acid sequence of the encoded protein. Encoding a secretory or transmembrane protein. D329
1 is a full-length clone, containing the entire coding sequence of a secreted protein (also referred to herein as "D3291 protein"). The nucleotide sequence of the 5 ′ portion of D3291 determined this time is shown in SEQ ID NO: 14
Shown in SEQ ID NO: 15 shows what the applicant considers to be the correct reading frame for the coding region. The deduced amino acid sequence of D3291 protein corresponding to the above nucleotide sequence is shown in SEQ ID NO: 15. D329 containing poly A tail
Additional nucleotide sequence from the 3 'portion of 1 is shown in SEQ ID NO: 16. EcoRI / Not obtainable from the deposit containing clone D3291
The I restriction fragment should be approximately 3100 bp in length. The nucleotide sequence of D3291 disclosed herein was searched against the GenBank and GeneSeq databases using BLASTA / BLASTX and FASTA search protocols. D329-1 is W46599 (zc32g12.r1 Soares senescent fibrobla
It showed at least some similarity to the sequence identified as sts NbHSF Homo sapiens cDNA). The deduced amino acid sequence disclosed herein for D329_1 is
A search was performed against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative D329_1 protein is U40941 (coded for by C. eleg)
ans cDNA (CEESB82F) showed at least some similarity to the sequence identified. Based on sequence similarity, the D3291 protein and each similar protein or peptide may share at least some activity.

Clone "H6983" The polynucleotide of the present invention has been identified as clone H6983. H6
983 describes a method that is selective for cDNAs encoding secreted proteins (US Pat.
No. 5,366,637), isolated from adult blood (phytohemagglutinin and phorbol myristate acetate and peripheral blood mononuclear cells treated with mixed lymphocyte reaction) cDNA library, or the amino acid sequence of the encoded protein. Based on computer analysis, it was identified as encoding a secreted or transmembrane protein. H6983 is a full-length clone, including the entire coding sequence of a secreted protein (also referred to herein as "H6983 protein"). The nucleotide sequence of the 5 ′ portion of H6983 determined this time is shown in SEQ ID NO: 17
Shown in The additional internal nucleotide sequence from H6983 determined this time is shown in SEQ ID NO: 18. What the applicant considers the correct reading frame and deduced amino acid sequence encoded by such an internal sequence is shown in SEQ ID NO: 19. An additional nucleotide sequence from the 3 'portion of H6983, including the poly A tail, is shown in SEQ ID NO: 20. EcoRI / Not obtainable from the deposit containing clone H6983
The I restriction fragment should be approximately 1400 bp in length. The nucleotide sequence of H6983 disclosed herein was searched against the GenBank and GeneSeq databases using BLASTA / BLASTX and FASTA search protocols. H6983 was prepared from N69846 (za67e04.s1 Homo sapiens cDNA clone 2).
97630 3 'similar to SW RT05_YEAST P33759 PROBABLE MITOCHONDRIAL 40S RIBO
SOMAL PROTEIN S5), W36396 (mb75e09.r1 Soares mouse p3NMF19.5 Mus musculu
s cDNA clone 335272 5 '), and W81295 (zd85h08.r1 Soares fetal heart Nb
It showed at least some similarity to the sequence identified as HH19W Homo sapiens cDNA clone 347487 5 '). The deduced amino acid sequence disclosed herein for H6983 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative H6983 protein is L20296 (homology
with a procaryotic 30S ribosomal protein S5 [Saccharomyces cerevisiae])
And at least some similarity to the sequence identified as Z69727 (unknown [Schizosaccharomyces pombe]). Based on sequence similarity, H69
The 83 proteins and each similar protein or peptide may share at least some activity. H6983 has sequence similarity to several other ribosomal proteins and to polynucleotides encoding ribosomal proteins or proteins similar to ribosomal proteins.

Clone "H9632 20" The polynucleotide of the present invention has been identified as clone "H9632-20" . H
96320 is a method for selectively encoding cDNA encoding secreted proteins (see US Pat. No. 5,536,637) in adult blood (phytohemagglutinin and phorbol myristate acetate and peripheral blood treated with a mixed lymphocyte reaction). Mononuclear cells) Isolated from cDNA libraries or identified as encoding secreted or transmembrane proteins based on computer analysis of the amino acid sequence of the encoded protein. H9632 20 is a full-length clone and contains the entire coding sequence of a secreted protein (also referred to herein as "H9620 protein"). The nucleotide sequence of H96320 determined this time is shown in SEQ ID NO: 21.
SEQ ID NO: 22 shows what the applicant considers to be the correct reading frame and deduced amino acid sequence of H96320 protein corresponding to the above nucleotide sequence. Amino acids 23 to 35 are a putative leader / signal sequence, with the predicted mature amino acid sequence starting at amino acid 36. Alternatively, amino acids 23 to 35 are a transmembrane domain. EcoRI / No obtainable from the deposit containing clone H96320
The tI restriction fragment should be approximately 1240 bp in length. The deduced amino acid sequence disclosed herein for H96320 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative H9620 protein is AA184698 (mt58f09.r1 Soares 2NbMT Mu
s musculus cDNA clone 634121 5 'similar to TR G285995 G285995 ORF, CMPLE
Showed at least some similarity to the sequence identified as TE CDS.).
The predicted amino acid sequence disclosed herein for H96320 was searched against the GenPept and GeneSeq amino acid sequence databases using the BLASTX search protocol. The putative H9632 20 protein is D13626 (KIAA0001 [Homo sapiens]), U33447 (pu
tative G-protein-coupled receptor [Homo sapiens]), and W04246 (Human
It showed at least some similarity to the sequence identified as G-protein coupled receptor GPR3). Based on sequence similarity, the H96320 protein and each similar protein or peptide may share at least some activity.

Deposit of clones Clone AY421 2, BV2782, C5441, CC33233
CC365 40, CG684, D3291, H6983 and H96.
320, based on the Budapest Treaty, was originally filed on August 22, 1996 as an original deposit.
It was deposited on the day with the American Type Culture Collection under accession number ATCC 98145, from which clones containing each polynucleotide can be obtained.
All restrictions on public use of the deposited material are set forth in 37 CFR. F. R. §1.808
Except for the provisions of (b), it will be canceled by grant of patent. Each clone was transfected into separate bacterial cells (E. coli) as this complex deposit. EcoRI / NotI digestion (5 ′ site, EcoRI; 3)
Site, NotI) to obtain fragments of appropriate size for such clones so that each clone can be removed from the deposited vector. Each clone was deposited in either the pED6 or pNotS vector shown in FIG. The pED6dpc2 vector (pED6) was derived from pED6dpc1 by inserting a new polylinker to facilitate cDNA cloning (Kaufman et al. (1991). NAR 19: 4485-4490). Deleting DHFR,
The pNOTs vector was transformed into pMT2 (Kaufman et al. 1989. Mol. Cell. Biol.) By inserting a new polylinker and inserting the M13 origin of replication into the ClaI site.
9: 1741-1750). In some cases, the deposited clones "flip" (ie, flipped) in the deposited isolate. In such cases, the cDNA insert can still be isolated by EcoRI and NotI digestion. However, in that case, c.
To place the DNA, NotI will create a 5 'site and EcoRI will create a 3' site. The cDNA may be expressed from the deposited vector. Bacterial cells containing individual clones can be obtained from the complex deposit as follows: An oligonucleotide probe or probe should be designed to a sequence known as a particular clone. This sequence can be derived from the sequences herein or a combination of those sequences. The oligonucleotide probes used to isolate each full-length clone are shown below, but they should be the most reliable in isolating the clone of interest.

Clone probe sequence AY421 2 SEQ ID NO: 23 BV278 2 SEQ ID NO: 24 C544 1 SEQ ID NO: 25 CC332 33 SEQ ID NO: 26 CC365 40 SEQ ID NO: 27 CG684 4 SEQ ID NO: 28 D329 1 SEQ ID NO: 29 H6983 SEQ ID NO: 30 H963 20 SEQ ID NO: 31

The sequences listed above include an N at position 2, which position is a biotinylated phosphoramidite residue (eg, biotin phosphoramidite (1-dimethoxytrityloxy-2-) rather than a nucleotide in a preferred probe / primer. (Obtained by use of (N-biotinyl-4-aminobutyl) -propyl-3-O- (2-cyanoethyl)-(N, N'-diisopropyl) -phosphoramidite (Glen Research catalog number 10-1953)). Occupied. Preferably, the design of the oligonucleotide probe should follow these parameters: (a) Should be designed to be the region of the sequence where the number of undefined bases (N), if any, is minimal; (B) Tm of about 80 ° C. (2 ° C. for A or T, 4 ° C. for G or C)
(Assume ° C). Preferably, oligonucleotides should be labeled with [gamma]-< ³² > P-ATP (specific activity 6000 Ci / mmole) using T4 polynucleotide kinase, using methods widely used for oligonucleotide labeling. Other labeling methods can be used. Preferably, unincorporated label should be removed by gel filtration or other established methods. It should be quantified by measuring the amount of radioactivity incorporated into the probe with a scintillation counter. Preferably,
The specific activity of the resulting probe should be about 4e + 6 dpm / pmole. Preferably, the bacterial culture containing the pool of full-length clones should be thawed and 100 μl of the stock used to inoculate a sterile culture flask containing 25 ml of sterile L broth containing 100 μg / ml ampicillin. Preferably, the culture should be grown at 37 ° C. to reach saturation, and preferably the saturated culture should be fresh L
Should be diluted with broth. Preferably, a portion of these dilutions are plated at 37 ° C. on solid bacterial culture medium containing L-broth containing 100 μg / ml ampicillin and 1.5% agar in a 150 mm petri dish. The dilution and volume at which 5000 well-defined and well-separated colonies will grow when grown overnight should be determined. Other known methods for obtaining well-defined and well-separated colonies can also be used. The colonies should then be transferred to nitrocellulose filters using standard colony hybridization techniques and lysed, denatured, and baked. Then, preferably, 6 × SSC containing 0.5% SDS, 100 μg / ml yeast RNA, and 10 mM EDTA (205.3 × 175.3 g stock).
NaCl / liter, 88.2 g sodium citrate, pH 7.0 with NaOH
(About 10 ml per 150 mm filter) at 65 ° C. for 1 hour with gentle stirring. The probe is then preferably added to the hybridization mix so that the concentration is greater than or equal to 1e + 6 dpm / ml. The filter is then preferably washed with 500 ml of 2X SSC / 0.5% SDS without stirring at room temperature, preferably afterwards with gentle shaking at room temperature with 500 ml of 2X SSC / 0. Wash with 1% SDS. 65 ° C., 30 minutes to 1 hour, 0.1 × SSC / 0
. A third wash with 5% SDS is optimal. The filter is then preferably dried and subjected to autoradiography for a sufficient time to visualize positives on the X-ray film. Other known hybridization methods can be used. Positive colonies are picked, grown in medium, and the plasmid DNA is then isolated using standard procedures. The clone can then be verified by restriction analysis, hybridization analysis or DNA sequencing.

A fragment of the protein of the present invention that can exhibit a biological activity is also included in the present invention. Protein fragments may be linear or, for example, HUSaragovi
, et al., Bio / Technology 10, 773-778 (1992) and RSMcDowell, et al., J
Amer. Chem. Soc. 114, 9245-9253 (1992) (both references are deemed to be described herein by reference) may be cyclized using known methods. . For many purposes, including increasing the number of valencies at the protein binding site,
Such a fragment may be fused to a carrier molecule along with the immunoglobulin.
For example, a protein fragment may be fused to the Fc portion of an immunoglobulin via a "linker". For the bivalent form of the protein, such a fusion is made to the F molecule of the IgG molecule.
This can be done for part c. Such fusions may be obtained using other immunoglobulin isotypes. For example, a protein-IgM fusion will yield a ten-valent form of a protein of the invention. The invention also provides the full length and mature forms of the disclosed proteins. The full length form of such a protein has been identified in the sequence listing by translation of the nucleotide sequence of the disclosed clone. Mature forms of such proteins may be obtained by expressing the disclosed full-length polynucleotides (preferably those deposited with the ATCC) in appropriate mammalian cells or other host cells. The sequence of the mature form of the protein may be determined from the amino acid sequence of the full-length form. The present invention also provides a gene corresponding to the cDNA sequence disclosed herein. "
A “corresponding gene” is a region of the genome that is transcribed to produce mRNA from which cDNA is derived and whose flanking regions (coding sequences, 5 ′ and 3 ′ non-coding sequences are required for regulated expression of such gene). Translation region), or spliced exons, introns, promoters, enhancers,
As well as silencer or suppressor elements. Corresponding genes can be isolated using the sequence information disclosed herein. Corresponding genes can be isolated using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence information to identify and / or amplify genes in a suitable genomic library or other source of genomic material. An "isolated gene" is a gene that has been separated from flanking coding sequences, if present, present in the genome of the organism from which the gene was isolated.

The chromosomal location corresponding to the polynucleotide sequences disclosed herein may be determined, for example, by hybridizing the appropriately labeled polynucleotide of the invention to the chromosome in situ. By identifying significantly similar nucleotide sequences in public databases, such as expressed sequence tags (ESTs) that have already been mapped to specific chromosomal locations, the corresponding chromosomal location of the disclosed polynucleotides may be determined. For at least some of the polynucleotide sequences disclosed herein, public database sequences having at least some similarity to the polynucleotides of the present invention are listed by database accession number. A search using the GenBank accession number of these public database sequences to identify UniGene clusters of overlapping sequences was performed at the National Center for Biotechnology I
This can be done on the internet site provided by nformation, the address of which is http://www.ncbi.nlm.nih.gov/UniGene/. Many UniGene clusters so identified have already been mapped to specific chromosomal locations.

An organism is provided that has enhanced, reduced, or altered expression of a gene corresponding to a polynucleotide sequence disclosed herein. Desirable changes in gene expression are achieved by using antisense polynucleotides or by using ribozymes that bind and / or cleave mRNA transcribed from the gene (Albert and Morris, 1994, Trends Pharmacol. Sci. 15 (7): 250-2
54; Lavarosky et al., 1997, Biochem.Mol. Med. 62 (1): 11-22; and Hampel,
1998, Prog. Nucleic Acid Res. Mol. Biol. 58: 1-39; all of which are herein incorporated by reference). A transgenic animal having a multicopy gene corresponding to a polynucleotide disclosed herein, preferably obtained by transforming cells with a genetic construct that is stably maintained in transformed cells and their progeny. A transgenic animal is provided. Also provided are transgenic animals having altered genetic control regions that increase or decrease gene expression levels or alter the temporal or spatial pattern of gene expression (see European Patent No. 0 649 464 B1; All of which are considered to be described herein by reference). In addition, an organism is provided wherein the gene corresponding to the polynucleotide sequence disclosed herein has been incompletely or completely inactivated, either by insertion of an external sequence or by deletion of all or part of the corresponding gene. By insertion, preferably by insertion after incorrect resection of the transposable element (Plasterk, 1992, Bioessays 14 (9): 629-633; Zwaal et al., 19).
93, Proc. Natl. Acad. Sci. USA 90 (16): 7431-7435; Clark et al., 1994, Pr.
USA 91 (2): 719-722; all of which are deemed to be described herein by reference), or by homologous recombination, preferably by positive / negative genetic selection. Homologous recombination (Mansour et a
l., 1988, Nature 336: 348-352; US 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 deemed herein to be incorporated by reference), incomplete or complete gene inactivation can be performed. These organisms with altered gene expression
Preferably they are eukaryotes, and more preferably mammals. Such organisms are useful for developing non-human models for studying diseases associated with the corresponding gene, and for developing assay systems for identifying molecules that interact with protein products from the corresponding gene. Where the protein of the invention is membrane-bound (eg, a receptor), the invention also provides soluble forms of such protein. In such a form, some or all of the intracellular and transmembrane domains of the protein are removed so that the protein is sufficiently secreted from the host in which it was expressed. The intracellular and transmembrane domains of the protein of the present invention can be identified by known methods for determining such domains from sequence information. For example, TopP
Predict the position of the transmembrane domain in the amino acid sequence using the redII computer program. The domain is represented by the central position of the transmembrane domain and has at least 10 transmembrane amino acids on either side of the reported central residue. The proteins and protein fragments of the present invention comprise at least 25% (more preferably at least 50%, most preferably at least 75%) of the amino acid sequence of the disclosed protein.
And has at least 60% sequence identity (more preferably at least 75% identity, most preferably at least 90%) to the disclosed proteins.
Or 95% identity). Sequence identity is determined by comparing the amino acid sequences of the proteins when the sequences are aligned to maximize overlap and identity while minimizing sequence gaps. It has at least 75% sequence identity (more preferably at least 85% identity, most preferably at least 95% identity) to any of the disclosed protein segments, preferably 8 or more ( Proteins or protein fragments containing segments comprising more preferably 20 or more, most preferably 30 or more) contiguous amino acids are also encompassed by the present invention.

Specifically, WU-BLAST (Washington University BLAST) version 2. 0 software may be used to determine sequence identity, which is a publicly available unrestricted WBI based on NCBI-BLAST version 1.4.
-Built based on BLAST (Altschul and Gish, 1996, Local ali
gnment statistics, Doolittle ed., Methods in Enzymology 266: 460-480; Al
tschul et al., 1990, Basic local alignment search tool, Journal of Molec
ular Biology 215: 403-410; Gish and States, 1993, Identification of prot
ein coding regions by database similarity search, Nature Genetics 3: 266
-272; Karlin and Altschul, 1993, Applications and statistics for multipl
e high-scoring segments in molecular sequences, Proc. Natl. Acad. Sci. U
SA 90: 5873-5877 (all of these references are hereby incorporated by reference)). WU-BLAST for some UNIX platforms
Ftp://blast.wustl.edu/blast/executab version 2.0 executable
You can download from les. In addition to several support programs, a complete set of search programs (BLASTP, BLASTN, BLASTX,
TBLASTN and TBLASTX) are provided at this site. WU
-BLAST 2.0 is copyrighted and must not be sold or redistributed in any form without the consent of the author, but may be used for non-business or research purposes. Can be used for Pair (BLASTP, BLAST
N, BLASTX, TBLASTN, and TBLASTX), the gap alignment routine (gapped alignment routine)
routines) are integrated into the database search itself, resulting in highly sensitive, highly selective and easily interpretable results. Optimally, gapping can be turned off in all these programs. The default penalty (Q) for the length gap is Q = 9 for BLASTP and Q = 10 for BLASTN, but 1 to 8, 9, 10, including zero. It may be changed to any integer value such as 11, 12 to 20, 21 to 50, and 51 to 100. Residue 1 to widen the gap
The default penalty (R) per unit is R = 2 for protein and BLASTP, and R = 10 for BLASTN, but 0, 1, 2, 3
, 4, 5, 6, 7, 8, 9, 10, 11, 12 to 20, 21 to 50, 5,
It may be changed to an integer value such as 1 to 100. Any combination of Q and R may be used to compare sequences side-by-side to maximize overlap and identity and minimize sequence gaps. The default amino acid comparison matrix is BLOS
Although UM62, other amino acid comparison matrices such as PAM can be used.

[0030] Species homologs of the disclosed polynucleotides and proteins are also provided by the invention. The term "species homolog" herein is a protein or polynucleotide that relates to a species different from the source of the protein or polynucleotide, but as determined by one of skill in the art,
Those having significant sequence similarity. Preferably, the polynucleotide species homolog has at least 60% (more preferably at least 75%, most preferably at least 90%) sequence identity to the particular polynucleotide, and the protein species homolog is At least 30% (more preferably at least 45%)
%, Most preferably at least 60%). Here, sequence identity is determined by comparing the nucleotide sequence of a polynucleotide or the amino acid sequence of a protein juxtaposed such that overlap and identity are maximized and sequence gaps are minimized. Species homologs may be isolated and identified by generating appropriate probes or primers from the sequences provided herein, and then screening for an appropriate nucleic acid source from the desired species. Preferably, the species homolog is isolated from a mammalian species. Most preferably, the species homolog is, for example, Pan troglodyte
s, Gorilla gorilla, Pongo pygmaeus, Hylobates concolor, Macaca mulatta,
Papio papio, Papio hamadryas, Cercopithecus aethiops, Cebus capucinus, A
otus trivirgatus, Sanguinus oedipus, Microcebus murinus, Mus musculus, R
attus norvegicus, Crisetulus griseus, Felis catus, Mustela vison, Canis
It has been isolated from specific mammals such as familiaris, Oryctolagus cuniculus, Bos taurus, Ovis aries, Sus scrofa and Equus caballus, and has been genetically mapped to identify the genes in one species. It is possible to identify the association of gene sequence order between genomic organization and genomic organization of related genes in another species (O'Brien and Seuanez, 1988, Ann. Rev. Genet. 22: 323). -351;
O'Brien et al., 1993, Nature Genetics 3: 103-112; Johansson et al., 199
5, Genomics 25: 682-690; Lyons et al., 1997, Nature Genetics 15: 47-56;
O'Brien et al., 1997, Trends in Genetics 13 (10): 393-399; Carver and Stu
bbs, 1997, Genome Research 7: 1123-1137, all of which are deemed to be described herein by reference).

The present invention also relates to allelic variants of the disclosed polynucleotides, ie, the spontaneous identification of isolated polynucleotides that encode proteins identical, homologous or related to the protein encoded by the disclosed polynucleotides. Form. Preferably, the allelic variant is at least 60% (for a particular polynucleotide)
More preferably at least 75%, most preferably at least 90%), wherein the sequence identity is such that the polynucleotides are juxtaposed such that overlap and identity are maximized and sequence gaps are minimized. By comparing the nucleotide sequences of Allelic variants may be isolated and identified by generating appropriate probes or primers from the sequences described herein and screening for an appropriate source of nucleic acid from an individual of the appropriate species. The present invention also encompasses polynucleotides having sequences complementary to the sequences of the polynucleotides disclosed herein.

[0032] The present invention also relates to polynucleotides that are capable of hybridizing to the polynucleotides described herein under conditions of reduced stringency, more preferably under stringent conditions, and most preferably under very stringent conditions. Also encompasses nucleotides. The following table shows examples of strictness conditions. The very strict conditions are, for example, at least as strict as the conditions A to F, and the strict conditions are, for example, at least as strict as the conditions GL, and the reduced conditions are, for example, For example, it is at least as strict as the conditions M to R.

[0033]

[Table 1] a): Hybrid length is the expected length for the hybridizing region (s) of the hybridizing polynucleotide. When hybridizing a polynucleotide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be the length of the hybridizing polynucleotide. When polynucleotides of known sequence hybridize, the length of the hybrid can be determined by juxtaposing the polynucleotide sequences and identifying the region (s) of optimal sequence complementarity. b): SSPE (1XS) in hybridization and wash buffer
SPE may be replaced by _{0.15M NaCl, 10mM NaH 2 PO 4} , and 1.25 mM E DTA, a pH7.4 is) the SSC (IX SSC is 0.15 M NaCl and 15mM sodium citrate). After completion of hybridization, washing is performed for 15 minutes. * T _B -T _R : The hybridization temperature for hybrids expected to be less than 50 base pairs in length should be 5-10 ° C below the melting temperature (T _m ) of the hybrid. T _m is determined by the following equation. For hybrids less than 18 base pairs in length, T _m (° C) = 2 (# of A + T bases) + 4 (# of G + C base
s). For hybrids 18 to 49 base pairs in length, T _m (° C.) = 81.5 + 16.6 (log ₁₀ [Na ⁺ ]) + 0.41 (% G + C) − (600 / N), where N is the hybrid And the sodium ion concentration in the hybridization buffer ([Na ⁺ ] for 1 × SSC = 0.165 M).

Further examples of stringency conditions for polynucleotide hybridization are described in Sambrook, J., EF Fritsch, and T. Maniatis, 1989, Molecular Cloning:
A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Ha
rbor, NY, chapters 9 and 11, and Current Protocols in Molecular Biology,
1995, FM Ausubel et al., Eds., John Wiley & Sons, Inc., sections 2.10
and 6.3-6.4 and are deemed to be incorporated herein by reference. Preferably, each such hybridizing polynucleotide is at least 25% of the polynucleotide of the invention to be hybridized.
(More preferably, at least 50%, most preferably at least 75%) and at least 60% sequence identity (more preferably at least 75%) to the polynucleotide of the invention to be hybridized. , Most preferably at least 90% or 95% identity). Sequence identity is determined by comparing the amino acid sequences of the proteins when the sequences are aligned to maximize overlap and identity while minimizing sequence gaps.

The isolated polynucleotide encoding the protein of the present invention is described in Kaufman et al., Nucl.
The protein may be recombinantly produced by operably linking it to an expression control sequence such as the pMT2 or pED expression vector disclosed in eic Acids Res. 19, 4485-4490 (1991). Many suitable expression control sequences are known in the art. Many suitable expression control sequences are known in the art. General methods for recombinant protein expression are also known and are described in R. Kaufman, Methods in Enzymology 185, 537-566.
(1990) is a typical example. "Operably linked" as defined herein means that the isolated polynucleotide of the present invention and the expression control sequence are placed in a vector or cell and transformed (transfected) with the ligated polynucleotide / expression control sequence. It is meant to be expressed by a host cell. Many types of cells can act as suitable host cells for expressing the proteins of the present invention.
Mammalian cells include, for example, monkey COS cells, Chinese hamster ovary (CH
O) obtained from in vitro culture of cells, human kidney 293 cells, human epidermal A431 cells, human Colo205 cells, 3T3 cells, CV-1 cells, other transformed primate cell lines, normal diploid cells, primary tissues Cell line, primary explant, H
Includes eLa cells, mouse cells, BHK, HL-60, U937HaK or Jurkat cells. Alternatively, the protein can be produced in lower eukaryotic cells such as yeast or prokaryotic cells such as bacteria. A potentially suitable yeast strain is Saccharomyces ce
revisiae, Schizosaccharomyces pombe, Kluyveromyces strain, Candida, or yeast strains capable of expressing heterologous proteins. A potentially suitable bacterial strain is Escherichia
E. coli, Bacillus subtilis, Salmonella typhimurium, or bacterial strains capable of expressing heterologous proteins. If the protein is produced in yeast or bacteria, it may be necessary to modify the protein produced therein, for example, by phosphorylation or glycosylation at appropriate sites to obtain a functional protein. Such covalent linkages may be made using known chemical or enzymatic methods. The isolated polynucleotides of the present invention may be operably linked to appropriate control sequences in one or more insect expression vectors and the protein obtained by using an insect expression system. Materials and methods for baculovirus / insect cell expression systems are described, for example, in In
Commercially available in kit form from Vitrogen, San Diego, California, USA (MaxBac ^R kit), such methods are well known in the art and are described in Summers and Smith, Texas Agricultural Experiment Station Bulletin No. 1555 (1987).
) (Assumed to be described herein by reference). As used herein, insect cells capable of expressing a polynucleotide of the present invention have been "transformed." The protein of the present invention may be produced by culturing the transformed host cell under culture conditions suitable for expressing the recombinant protein. The resulting expressed protein may then be purified from such culture (ie, medium or cell extract) using known purification processes, such as gel filtration and ion exchange chromatography. Purification of the protein may also be accomplished by affinity columns containing an agent that will bind to the protein; one or more column steps with an affinity column such as Concanavalin A-agarose, Heparin-Toyopearl ^R or Cibacrom blue 3GA Sepharose ^R ; One or more steps using hydrophobic interaction chromatography with a resin such as phenyl ether, butyl ether, or propyl ether; or immunoaffinity chromatography. Alternatively, the protein of the invention may be expressed in an easily purified form. For example, maltose binding protein (MBP), glutathione-S-tonsferase (GST)
Alternatively, it may be expressed as a fusion protein such as a fusion protein with thioredoxin (TRX). Kits for expression and purification of such fusion proteins are available from New En
Commercially available from glan BioLab (Beverly, MA), Pharmacia (Piscataway, NJ) and InVitrogen. The protein can also be tagged with an epitope and then purified by using a specific antibody directed against such epitope. One such epitope ("flag") is commercially available from Kodak (New Haeven, CT). Finally, the protein is further purified using one or more reverse phase high quality liquid chromatography (RP-PLC) steps using a hydrophobic RP-HPLC medium, such as silica gel with pendant methyl or other aliphatic groups. can do. Some or all of the above purification steps may be used in various combinations to obtain a substantially homogeneous isolated recombinant protein. The protein thus purified is substantially free of other mammalian proteins and is defined in the present invention as "isolated protein".

The protein of the present invention may be used as a product of a transgenic animal, for example, a component of milk of a transgenic bovine, goat, pig, or sheep characterized by somatic cells or germ cells containing a nucleotide sequence encoding the protein. May be expressed as The protein may be produced by known conventional chemical synthesis. Methods for constructing the protein of the present invention by synthetic means are known to those skilled in the art. Synthetically constructed protein sequences may have common biological properties, including protein activity, by sharing primary, secondary or tertiary structure and / or conformational properties. Thus, they may be used as biological activities or immunological substitutes for naturally occurring purified proteins in immunological processes for screening therapeutic compounds and generating antibodies.

The proteins described herein are characterized by an amino acid sequence similar to the amino acid sequence of the purified protein, but may be modified therein, either naturally or by derivatization. For example, modifications of the peptide or DNA sequence can be made by those skilled in the art using known methods. Modifications of interest in the protein sequence include changes, substitutions, exchanges, insertions or deletions of selected amino acid residues in the coding sequence. For example, one or more cysteine residues may be deleted or replaced with another amino acid to alter the conformation of the molecule. Methods for such changes, substitutions, exchanges, insertions or deletions are well known to those skilled in the art (eg, US Pat. No. 415).
No. 8584). Preferably, such changes, substitutions, exchanges, insertions or deletions retain the desired activity of the protein. Other fragments and derivatives of the protein sequence that are expected to retain protein activity in whole or in part, and thus may be useful in screening or other immunological methods, can also be made by those skilled in the art from the disclosure herein. . Such modifications are believed to be encompassed by the present invention.

Uses and Biological Activities The polynucleotides and proteins of the present invention are believed to exhibit one or more of the following uses or biological activities, including those associated with the assays cited herein. Can be The uses or activities described with respect to the proteins of the present invention may be due to the administration or use of such proteins, or the administration or use of polynucleotides encoding such proteins (eg, such as a vector suitable for gene therapy or DNA transfer). Can be provided.

Research Uses and Utility The polynucleotides provided by the present invention can be used for various purposes by a research population. For analysis, characterization or therapeutic use, as a marker of tissue in which the corresponding protein is preferentially expressed (constitutively or at a particular stage of tissue differentiation or development or in a disease state), and To identify potential genetic diseases as compared to the patient's endogenous DNA, as chromosomal markers or tags (if labeled) for identification of chromosomes or mapping of relevant gene locations, as molecular weight markers for Southern gels The use of known sequences in the process of discovering other novel polynucleotides as a source for obtaining PCR primers for genetic fingerprinting to find new related DNA sequences to use as hybridization probes "Gene chips" or other probes as probes for subtraction To select and generate oligomers for binding to carriers, to test expression patterns, to generate anti-protein antibodies using DNA immunization techniques, and to generate anti-DNA antibodies or to generate additional immunizations. Polynucleotides can be used as antigens to induce a response. If encoding a protein that binds or potentially binds to another protein (e.g., as in a receptor-ligand interaction), identify the other protein that binds, and / or an inhibitor of the binding interaction Polynucleotides can be used in an interaction trap assay (eg, as described in Gyuris et al., Cell 75: 791-803 (1993)) to identify The proteins provided by the present invention may be used in assays to determine biological activity, including a group of multiple proteins for high-throughput screening; to generate antibodies or induce other immune responses; As a reagent (including a labeling reagent) in an assay designed to quantitatively determine the level of a protein (or its receptor) in a biological fluid; the corresponding protein is preferentially expressed (constitutively Or expressed at a particular stage of tissue differentiation or development, or in a disease state) as a marker for tissue; and, of course, may be used to isolate relevant receptors or ligands. Where a protein binds or potentially binds to another protein, the protein can be used to identify other proteins that bind, or to identify inhibitors of the binding interaction. Using proteins involved in these binding interactions, screening for peptides or small molecular inhibitors or agonists for the binding interaction can also be performed. Any or all of these research applications can be evolved into reagent grade or kit formats for commercialization as research products. Methods for performing the above uses are well known to those skilled in the art. A literature disclosing such a method can be found in "Molecular Cloning: A Laboratory Manual", 2nd ed., Cold Sp.
ring Harbor Laboratory Press, Sambrook, J., EF Fritsch and T. Maniatis
eds., 1989, and "Methods in Enzymology: Guide to Molecular Cloning Tec
hniques ", Academic Press, Berger, SL and AR Kimmel eds., 1987.

Use as Nutrition The polynucleotides and proteins of the present invention can be used as nutrient sources or additives. Such uses include, but are not limited to, use as a protein or amino acid additive, use as a carbon source, use as a nitrogen source, and use as a carbohydrate source. In such cases, the proteins or polynucleotides of the present invention may be added as food for a particular organism, or may be administered as a separate individual or liquid formulation, such as in the form of a powder, pill, solution, suspension or capsule. it can. In the case of a microorganism, the protein or polynucleotide of the present invention can be added to a medium in which the microorganism is cultured.

Cytokines and Cell Proliferation / Differentiation Activity The protein of the present invention may exhibit cytokine activity, cell proliferation activity (induction or inhibition) or cell differentiation activity (induction or inhibition), or produce other cytokines in certain cell populations. Can be induced. Many proteinaceous factors (including all known cytokines) that have been found to date have demonstrated activity in one or more factor-dependent cell proliferation assays, thus making the assay a convenient method of confirming cytokine activity. Serve as. The activity of the protein of the present invention is determined in cell lines (32D, DA
2, DA1G, T10, B9, B9 / 11, BaF3, MC9 / G, M + (pr
eB M +), 2E8, RB5, DA1, 123, T1165, HT2, CTL L2, TF-1, Mo7e and CMK). Confirmed by either of The activity of the protein of the invention may be measured, inter alia, by the following method: Assays for T cell or thymocyte proliferation are described in Current Protocols in Immunology, Ed by JE Coligan, AM Kruisbeek, D .;
H. Margulies, EM 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: 34943500, 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, but are not limited thereto. Assays for cytokine production and / or proliferation of spleen cells, lymph node cells or thymocytes are described in Polyclonal T cell stimulation, Kruisbeek, AM and Shevach, EM In Cur
rent Protocols in Immunology. JEea Coligan eds. Vol 1 pp. 3.12.1-3.1
2.14, John Wiley and Sons, Toronto. 1994; and Measurement of mouse and h
uman Interferon g, Schreiber, RD In Current Protocols in Immunology.
JEea Coligan eds.Vol 1 pp. 6.8.1-6.8.8, John Wiley and Sons, Toront
o. Includes, but is not limited to, those described in 1994. Assays for proliferation and differentiation of hematopoietic and lymphogenic cells are described in the Measurement of Human and Murine Interleukin 2 and Interleukin 4, Bottoml
y, K., Davis, LS and Lipsky, PE In Current Protocols in Immunology.
JEea Coligan eds.Vol 1 pp. 6.3.1-6.3.12, John Wiley and Sons, Toron
to. 1991; deVries et al., J. Exp.Med. 173: 1205-1211, 1991; Moreau et al.
., Nature 336: 690-692, 1988; Greenberger et al., Proc. Natl. Acad. Sci.
USA 80: 2931-2938, 1983; Measurement of mouse and human interleukin 6
-Nordan, R. In Current Protocols in Immunology. JEea Coligan eds. V
ol 1 pp. 6.6.1-6.6.5, John Wiley and Sons, Toronto.1991; Smith et al.,
Proc. Natl. Acad. Sci. USA 83: 1857-1861, 1986; Measurement of human I
nterleukin 11-Bennett, F., Giannotti, J., Clark, SC and Turner, K. J
. In Current Protocols in Immunology. JEea Coligan eds. Vol 1 pp.
6.15.1 John Wiley and Sons, Toronto. 1991; Measurement of mouse and hum
an Interleukin 9-Ciarletta, A., Giannotti, J., Clark, SC and Turner,
KJ In Current Protocols in Immunology. JEea Coligan eds. Vol 1 p
p. 6.13.1, including, but not limited to, those described in John Wiley and Sons, Toronto. 1991. Assays for the response of T cell clones to antigens, specifically identifying APC-T cell interactions and direct T cell effects by measuring proliferation and cytokine production, are described in Current Protocols in Immunology, Ed by JE. Coligan, AM Kruisbeek, D.
H. Margulies, EM 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. Sc
i. 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, including, but not limited to.

Immunostimulatory or Suppressive Activity The protein of the present invention may also exhibit immunostimulatory or immunosuppressive activity, including but not limited to the activity in the assays described herein. Protein is
It may be useful in a variety of deficiencies and disorders, including severe immunodeficiency complications (SCID), for example, in up-regulating or down-regulating T and / or B lymphocyte proliferation, and in N
It may be useful in activating the cytolytic activity of K cells and other cell populations. These immunodeficiencies can be genetic and can include viral (
For example, it may be caused by HIV) and bacterial or fungal infections, or may be caused by an autoimmune disease. More specifically, infectious diseases caused by viruses, bacteria, fungi or other infectious agents, for example, various fungal infections such as HIV, hepatitis virus, herpes virus, mycobacteria, Leishmania, malaria and Candida species, It can be treated using the protein of the present invention. Of course, in this regard, the proteins of the present invention may be used where a boost to the immune system is indicated, ie, in the treatment of cancer. Autoimmune diseases that may be treated using the protein of the present invention include, for example, multiple sclerosis,
Systemic deep elitomades, rheumatoid arthritis, autoimmune pneumonia, Guillain-Barre syndrome, autoimmune thyroiditis, insulin-dependent diabetes mellitus, myasthenia gravis, graft-versus-host disease and autoimmune inflammatory Eye diseases. Such proteins of the invention may be used in the treatment of allergic reactions and conditions, such as asthma or other respiratory diseases. Other conditions in which immunosuppression is desired, including, for example, asthma (particularly allergic asthma) and related respiratory problems, may be treated with the proteins of the invention. Other conditions for which immunosuppression is desired (including, for example, organ transplantation)
May be treated using the protein of the present invention. The proteins of the present invention can also be used to enable an immune response 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 interfering with the induction of an immune response. The function of activated T cells may be inhibited by suppressing T cell responses, or by inducing specific resistance in T cells, or both. In general, immunosuppression of a T cell response is an active, non-antigen-specific process that requires continuous exposure of T cells to an inhibitor. Tolerance means non-responsiveness or anergy in T cells, and differs from immunosuppression in that it is generally antigen-specific and persists after exposure to the tolerizing agent is stopped.
Operationally, resistance may be indicated by a lack of a T cell response when exposed to a specific antigen in the absence of a tolerizing agent. Down-regulating or preventing one or more antigen functions, including but not limited to B-lymphocyte antigen functions (eg, such as B7), eg, high levels of lymphokines by activated T cells Interference with synthesis may be useful in tissue, skin and organ transplantation and graft versus host disease (GVHD). For example, blocking T cell function should reduce tissue destruction in tissue transplantation. Typically, in tissue transplants, rejection of the graft is initiated by the recognition of the tissue piece as foreign by T cells, followed by an immune response that destroys the graft. A monomeric form having the activity of a molecule that inhibits or blocks the interaction of a B7 lymphocyte antigen with its native ligand on immune cells (another B lymphocyte antigen (eg, B7-1, B7-3)) Mixed with peptides,
Alternatively, pre-transplant administration of a soluble, monomeric form of a peptide with B7-2 activity alone, induces binding of the molecule to its native ligand on immune cells without transmitting a responsive costimulatory signal. there is a possibility. Blocking B lymphocyte antigen function in this regard prevents cytokine synthesis by immune cells, such as T cells, and thus acts as an immunosuppressant. Moreover, the lack of costimulation may be sufficient to cause a loss of T cell response. The induction of long-term tolerance by B lymphocyte antigen blocking reagents may obviate the need for repeated administration of these blocking reagents. To achieve sufficient immunosuppression or resistance in a subject, it may also be necessary to block the function of the B lymphocyte antigen combination. The efficacy of individual blocking reagents in preventing organ transplant rejection or GVHD can be evaluated using animal models that can predict efficacy in humans. Examples of suitable systems that can be used include allogeneic heart grafts in rats and xenogeneic pancreatic islet cell grafts in mice, both of which are used to test the immunosuppressive effects of CTLA4Ig fusion proteins in vivo. (Lensch
ow et al., Science 257: 789-792 (1992) and Turka et al., Proc Natl. Acad.
Sci. USA, 89: 11102-11105 (1992)). In addition, a rat model of GVHD (Paul ed., Fundamental Immunology, Ravan Press, New York, 1
989, pp. 846-847) can be used to determine the blocking effect of B lymphocyte antigen function on the progression of the disease in vivo. Also, blocking antigen function can be therapeutically useful for treating autoimmune diseases. Many autoimmune diseases are the result of inappropriate activation of T cells that are reactive to self tissue and promote the production of cytokines and autoantibodies that are involved in the pathology of the disease. Interfering with the activation of autoreactive T cells may reduce or eliminate the symptoms of the disease. Receptors for B lymphocyte antigens: autoantibodies or T cell-derived that inhibit T cell activation using administration of reagents that block T cell costimulation by disrupting ligand interactions and may be involved in the disease process Can interfere with the production of cytokines. In addition, blocking reagents can induce antigen-specific resistance of autoreactive T cells that can lead to long-term remission of the disease. The effectiveness of a blocking reagent in preventing or ameliorating an autoimmune disease can be determined using a number of well-characterized animal models for human autoimmune diseases. Examples include murine experimental autoimmune encephalitis, systemic deep erythematosus in MRL lpr / lpr or NZB hybrid mice, murine autoimmune collagen arthritis, diabetes in NOD mice and BB rats, and experimental murine myasthenia gravis. (Paul ed., Fundamental Immunology, Raven Pres
s, New York, 1989, pp. 840-856). Up-regulation of antigen function (preferably B-lymphocyte antigen function) as a means to up-regulate the immune response is also therapeutically useful.
Up-regulation of the immune response may be in the form of promoting an existing immune response or eliminating the initial immune response. For example, enhancing an immune response by stimulating B lymphocyte antigen function may be useful in the case of a viral infection.
In addition, systemic viral diseases such as influenza, common cold, and encephalitis may be ameliorated by systemic administration of stimulatory forms of B lymphocyte antigen.

Alternatively, T cells are taken from a patient and co-stimulated in vitro with a viral antigen to which ACPs expressing the peptide of the invention have been added, or combined with a stimulatory form of a soluble peptide of the invention, The T-cells activated in vitro may then be reintroduced into the patient to enhance the anti-viral immune response in the infected patient. Another method of promoting an antiviral immune response is
The infected cells are removed from the patient and transfected with them a nucleic acid encoding a protein of the invention as described herein so that the cells express all or part of the protein on their surface. Will be re-introduced. The infected cells would then be able to transmit the costimulatory signal to the T cells in vivo and thereby activate the T cells. In another application, up-regulation or promotion of antigen function (preferably B lymphocyte antigen function) may be useful in inducing tumor immunity. Administering a tumor cell (eg, sarcoma, melanoma, lymphoma, leukemia, neuroblastoma, carcinoma) transfected with a nucleic acid encoding at least one peptide of the present invention to a tumor-specific tumor in the subject Can overcome resistance. If desired, tumor cells can be transfected to express the peptide combination. For example, an expression vector that directs the expression of tumor cells obtained from a patient in combination with a peptide having B7-2-like activity alone or with a peptide having B7-1-like activity and / or B7-3-like activity, It can be transfected ex vivo. The transfected tumor cells are returned to the patient and the peptide is expressed on the surface of the transfected cells. Alternatively, tumor cells can be targeted for transfection in vivo using gene therapy. The presence of a peptide of the invention having the activity of a B lymphocyte antigen on the surface of tumor cells provides the necessary costimulatory signals for T cells to induce an immune response to transfected tumor cells mediated by T cells I do. In addition, MHC
Tumor cells lacking class I or MHC class II molecules, or sufficient MHC
Class I or tumor cells can not be re-expressed MHC class II molecules, MHC class Iα chain protein and beta ₂ microglobulin protein or an MHC class IIα chain Contact and MHC class IIβ chain all or part of the protein (e.g., truncations protein The nucleic acid encoding the cytoplasmic domain can be transfected, thereby allowing the expression of class I or class II MHC proteins on the cell surface. Expression of appropriate Class I or Class II HMC in combination with a peptide having the activity of a B lymphocyte antigen (eg, B7-1, B7-2, B7-3) is transfected by T cells. Induces an immune response against tumor cells. If desired, a gene encoding an antisense construct that blocks expression of an MHC class II binding protein, such as an invariant chain, can be co-transfected with a DNA encoding a peptide having B lymphocyte antigen activity. It can also enhance the presentation of tumor-associated antigens and induce tumor-specific immunity. Thus, induction of an immune response mediated by T cells in a human subject may be sufficient to overcome tumor-specific resistance in the subject. The activity of the protein of the invention may be measured, inter alia, by the following method: Assays suitable for thymocyte or spleen cell cytotoxicity are described in Current Protocols in Immunology, Ed by JE Coligan, AM Kruisbeek, D .;
H. Margulies, EM 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
Natl. Acad. Sci. USA 78: 2488-2492, 1981; Herrmann et al., Proc.
J. Immunol. 128: 1968-1974, 1982; Handa et al., J. Immunol. 135: 1564-1
572, 1985; Takai et al., J. Immunol. 137: 3494-3500, 1986; Takai et al.,
J. Immunol. 140: 508-512, 1988; Herrmann et al., Proc. Natl. Acad. Sci.
USA 78: 2488-2492, 1981; Herrmann et al., J. Immunol. 128: 1968-1974, 1
982; Handa et al., J. Immunol. 135: 1564-1572, 1985; Takai et al., J. Im.
munol. 137: 3494-3500, 1986; Bowmanet al., J. Virology 61: 1992-1998; Ta
kai 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, including, but not limited to. Assays for T cell-dependent immunoglobulin responses and isotype switching, specifically to modulate T cell-dependent antibody responses and identify proteins that affect Th1 / Th2 profiles, are described in Maliazewski, J. Immunol. 144: 3028-3033, 1990, including but not limited to assays for B cell function as described in In vitro antibody production, Mond, JJ and Brunswick, M. In Current.
Protocols in Immunology JEColigan eds.Val 1 pp.3.8.1-3.8.16, John Wile
y and Sons, Tronto 1994. The mixed lymphocyte reaction (MLR) assay, which identifies proteins that primarily produce Th1 and CTL responses, is described in Current Protocols in Immunology, Ed by JE Coligan, AM Kruisbeek, D .;
H. Margulies, EM 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: 50
8-512, 1988; including but not limited to the assays described in Bertagnolli et al., J. Immunol. 149: 3778-3783, 1992. Dendritic cell-dependent assays, specifically identifying proteins expressed by dendritic cells that activate intact T cells, are described by Guery et al., J. Immunol. 134: 536-544, 1995; Inaba. et al., Journal of E
xperimental Medicine 173: 549-559, 1991; Macatonia et al., Journal of Im
munology 154: 5071-5079, 1995; Porgador et al., Journal of Experimental.
Medicine 182: 255-260, 1995; Nair et al., Journal of Virology 67: 4062-4
069, 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., J
Includes, but is not limited to, the assays described in ournal of Experimental Medicine 172: 631-640, 1990. Assays for lymphocyte survival / apoptosis, specifically identifying proteins that prevent apoptosis following superantibody induction, as well as proteins that regulate lymphocyte homeostasis, are described by Darzynkiewicz et al., Cytometry 13: 795-808, 1992; Gorczyca et al.
, Leukemia 7: 659-670, 1993; Gorczyca et al., Cancer Research 53: 1945-1
951, 1993; Itoh et al., Cell 66: 233-243, 1991; Zacharchuk, Journal of I
mmunology 145: 4037-4045, 1990; Zamai et al., Cytometry 14: 891-897, 199
3; including but not limited to the assays described in Gorczyca et al., International Journal of Oncology 1: 639-648, 1992. Early stage T cell commitment and development assays
Antica et al., Blood 84: 111-117, 1994; Fine et al., Cellular Immunology 15
5: 111-122, 1994; Galy et al., Blood 85: 2770-2778, 1995; Toki et al., Pr
USA 88: 7548-7551, 1991, including but not limited to oc. Natl. Acad. Sci.

Hematopoietic modulatory activity The proteins of the present invention are useful in regulating hematopoiesis and are therefore useful in treating bone marrow and lymphocyte deficiencies. Even minimal biological activity supporting colony forming cells or factor-dependent cell lines has been implicated in regulating hematopoiesis.
For example, supporting the proliferation of erythroid progenitor cells alone, or in combination with other cytokines, for example, showing usefulness in the treatment of various anemias, or in combination with radiation therapy / chemotherapy, erythroid progenitor cells And / or stimulating the production of erythroblasts; supporting the proliferation of bone marrow cells such as granulocytes and monocytes / macrophages (ie, traditional CSF activity), eg, in combination with chemotherapy, Useful in preventing myelosuppression that occurs; supporting proliferation of megakaryocytes and then platelets, and thereby preventing or treating various platelet disorders such as thrombocytopenia; It is also commonly used instead of or in complement to platelet infusion; and / or hematopoietic stem cells (mature Becomes all of hematopoietic cells, therefore, a variety of stem cell disease (
It is usually a disease that is treated by transplantation, and is useful in supporting the growth of, for example, aplastic anemia and paroxysmal hemoglobinuria), and is also useful in vivo or ex vivo. It is also useful in repopulating the stem cell compartment as normal cells after radiation / chemotherapy (ie, in combination with bone marrow transplantation) or after being engineered for gene therapy. In particular, the activity of the protein of the present invention may be measured by the following method. Assays suitable for the proliferation and differentiation of various hematopoietic cell lines have already been cited. Assays for embryonic stem cell differentiation, particularly those that identify proteins that affect embryonic hematopoiesis, are described in Johansson et al. Cellular Biology 15: 141-151, 1995; Keller et al.
, Molecular and Cellular Biology 13: 473-486, 1993; McClanahan et al., B
lood 81: 2903-2915, 1993, including, but not limited to. Assays for stem cell survival and differentiation, particularly identifying proteins that modulate lympho-hematopoiesis, are described in Methylcellulose colony forming assays, Freshney, MG In Culture of Hema.
topoietic Cells.RI 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 h
igh proliferative potential, McNiece, IK and Briddell, RA In Culture
of Hematopoietic Cells.RI Freshney, et al. eds.Vol pp. 23-39, Wile
y-Liss, Inc., New York, NY. 1994; Neben et al., Experimental Hematology.
22: 353-359, 1994; Cobblestone area forming cell assay, Ploemacher, RE
In Culture of Hematopoietic Cells.RI Freshney, et al. Eds.Vol pp.
1-21, Wiley-Liss, Inc., New York, NY. 1994; Long term bone marrow cultu
res in the presence of stromal cells, Spooncer, E., Dexter, M. and Allen
, T. In Culture of Hematopoietic Cells.RI Freshney, et al. Eds.Vol.
pp. 163-179, Wiley-Liss, Inc., New York, NY. 1994; Long term culture ini
tiating cell assay, Sutherland, HJ In Culture of Hematopoietic Cells.
RI Freshney, et al. Eds.Vol pp. 139-162, Wiley-Liss, Inc., New York,
Includes, but is not limited to, the assays described in NY. 1994.

Tissue Proliferation Activity The protein of the present invention is useful in compositions used for the growth or regeneration of bone, cartilage, key, ligament and / or nervous tissue, as well as for wound healing and tissue repair, and in the treatment of burns, lacerations and ulcers. Has utility. The protein of the present invention, which induces cartilage and / or bone growth in an environment where bone is not normally formed, finds use in healing fractures and cartilage damage or defects in humans and other animals. Such formulations using the protein of the present invention are useful for reducing closed and open fractures and for improving fixation of artificial joints. De novo osteogenesis induced by osteogenic agents contributes to the repair of congenital, traumatic or oncological resection-induced craniofacial defects and is also useful in cosmetic surgery. The protein of the present invention may be used in the treatment of periodontal disease and other tooth restoration processes. Such agents provide an environment for attracting osteogenic cells, stimulate the growth of osteogenic cells,
Alternatively, it can induce differentiation of osteogenic cell precursors. The protein of the present invention may be used to stimulate osteoporosis or osteoarthritis, for example, by stimulating bone and / or cartilage repair or by blocking the process of tissue destruction mediated by inflammation or inflammatory processes (collagenase activity, osteoclast activity, etc.). May also be useful in the treatment of Another category of tissue development activity that may be due to the proteins of the present invention is key / ligament formation. Proteins of the invention that induce the formation of key / ligament-like or other tissues in an environment in which such tissues are not normally formed can be used to prevent key or ligament laceration, deformation and other key or ligament defects in humans and other animals. Applied to healing, Such formulations using the key / ligament-like tissue inducing protein may be used to prevent damage to the key or ligament tissue as well as to improve the fixation of the key or ligament to bone or other tissue. The de novo key / ligament-like tissue formation induced by the compositions of the present invention contributes to the repair of congenital, traumatic, or oncological resection-induced craniofacial defects, and furthermore the attachment of keys or ligaments Or it is also useful in cosmetic surgery for repair. The compositions of the present invention provide an environment for attracting key- or ligament-forming cells, stimulate the growth of key- or ligament-forming cells, and induce the differentiation of key- or ligament-forming cell precursors. Or induce the growth of key / ligament cells or progenitors ex vivo so as to effect tissue repair when returned to vivo.
The compositions of the present invention are also useful for keratitis, carpal tunnel syndrome and other key or ligament defects. The compositions of the present invention may include a suitable matrix and / or sequestering agent, such as a carrier well known in the art. The protein of the present invention is useful for the proliferation of nerve cells and the regeneration of nerve and brain tissues, namely, central and peripheral nervous system diseases and neuropathies and mechanical diseases and traumatic diseases (including degeneration, death or trauma to nerve cells or nerve tissues). ) May also be useful. More particularly, diseases of the peripheral nervous system such as peripheral nerve injury, peripheral and localized neuropathy, and Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral spinal sclerosis and Shy-Drager
The protein may be used in the treatment of diseases of the central nervous system, such as the syndrome. Additional conditions that may be treated according to the present invention include mechanical and traumatic diseases such as spinal cord diseases, cerebrovascular diseases such as head trauma and stroke. Peripheral neuropathy caused by chemotherapy or other medical therapies can be treated with the proteins of the present invention. The protein of the present invention is also useful for promoting more preferable and prompt closure of non-healing wounds including (but not limited to) pressure ulcers, ulcers related to vascular dysfunction, surgical and trauma wounds and the like. It is possible. The protein of the present invention may also be used in other tissues such as organs (including pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth muscle, skeletal muscle or cardiac muscle), and vascular (including vascular endothelium) tissue. Or an activity for promoting the growth of cells constituting such a tissue. Part of the desired effect may be the regeneration of normal tissue by suppressing fibrotic scarring. The proteins of the present invention may also exhibit angiogenic activity. The proteins of the present invention may also be useful in protecting or regenerating intestines and treating conditions caused by lung or liver fibrosis, reperfusion injury of various tissues, and systemic cytokine damage. The protein of the present invention promotes or suppresses differentiation of the above-mentioned tissue from precursor tissue or cells;
Alternatively, it may be useful for suppressing the growth of the tissue. The activity of the protein of the invention may be measured, inter alia, by the following method: Tissue regeneration activity assay is described in WO 95/16035 (bone, cartilage, key).
International Publication WO95 / 05846 (nerves, neurons); International Publication WO91 / 05
7491 (skin, endothelium), but is not limited thereto. Wound healing activity assays are described in Winter, Epidermal Wound Healing, pps. 71-112.
(Maibach, HI and Rovee, DT, eds.), Year Book Medical Publishers, Inc.,
Including, but not limited to, those described in Chicago (modified by Eaglstein and Mertz, J. Invest. Dermatol 71: 382-384 (1978)).

Activin / Inhibin Activity The protein of the present invention may also exhibit activin- or inhibin-related activity. Inhibins are characterized by their ability to inhibit follicle stimulating hormone (FSH) release,
Activins are characterized by their ability to stimulate the release of follicle stimulating hormone (FSH). Thus, the protein of the present invention, alone or in the form of a heterodimer with a member of the inhibin α family, reduces the fertility of female mammals and reduces the spermatogenesis of male mammals. Can be useful as Administration of a sufficient amount of another inhibin can induce infertility in these mammals. Alternatively, the protein of the present invention, as a homodimer or as a heterodimer with subunits of other proteins of the inhibin-β group, can be used to stimulate fertility based on the ability of activin molecules to stimulate FSH release from anterior pituitary cells. May be useful as therapeutic agents that induce See, for example, U.S. Pat. No. 4,798,885. The proteins of the invention may also be useful for improving reproduction in sexually immature mammals, resulting in increased fertility during the life of livestock such as cattle, sheep and pigs. The activity of the protein of the invention may be measured, inter alia, by the following method: Activin / inhibin activity assays are described in Vale et al., Endocrinology 91:
562-572, 1972; Ling et al., Nature 321: 779-782, 1986; Vale et al., Natu
re 321: 776-779, 1986; Mason et al., Nature 318: 659-663,1985; Forage et
Natl. Acad. Sci. USA 83: 3091-3095, 1986, including but not limited to the assays described in US Pat.

Chemotaxis / chemokinesis activity The protein of the present invention may be used for the chemotaxis or chemokinesis activity of mammalian cells such as monocytes, neutrophils, T cells, mast cells, eosinophils and / or endothelial cells (for example, (Acting as a chemokine). Chemotactic and chemokinetic proteins can be used to recruit or attract a desired cell population to a desired site of action. Chemotaxis or chemokinesis proteins are particularly advantageous for treating injuries and other trauma to tissues and for treating local infections. For example, attracting lymphocytes, monocytes or neutrophils to a tumor or site of infection can improve the immune response to the tumor or infectious agent. Certain proteins or peptides have chemotactic activity on a particular cell population and, when stimulable, direct or indirectly direct the movement of such cell population. Preferably, the protein or peptide has the ability to directly stimulate the directed movement of the cell. Whether a particular protein has chemotactic activity on a cell population can be readily determined by using such a protein or peptide in a known assay for cell chemotaxis. The activity of the protein of the invention may be measured, inter alia, by the following method: Assays for chemotactic activity (assays to identify proteins that induce or interfere with chemotaxis) induce migration across cell membranes. Assays that measure the ability of a protein to induce adhesion of one cell population to another cell population as well as the ability of the protein to adhere to another cell population. Suitable assays for migration and adhesion are described in Current Protocols in Immunology, Ed by JE Coligan, AM Kruisbeek, DH
Margulies, EM Shevach, W. Strober, Pub. Greene Publishing Associates
and Wiley-Interscience (Chapter 6.12, Measurement of alpha and beta Chem
okines 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, including but not limited to the assays described.

Hemostatic and thrombolytic activity The proteins of the present invention may also exhibit hemostatic or thrombolytic activity. As a result, such proteins are useful in the treatment of various clotting disorders, including hereditary disorders such as hemophilia, or in the treatment of injuries caused by trauma, surgery, or other causes, as well as clotting and other hemostasis. Can be promoted. The protein of the present invention may be useful for the treatment and prevention of thrombolysis or inhibition of thrombus formation and conditions resulting therefrom (eg, myocardial infarction and infarction of central nervous system blood vessels (eg, stroke)). The activity of the protein of the invention may be measured, inter alia, by the following method: Assays for haemostatic and thrombolytic activities are described in Linet et al., J. Clin. Pharmacol. 26: 131-140,1986; Burdick et al. , Thro
mbosis Res. 45: 413-419, 1987; Humphrey et al., Fibrinolysis 5: 71-79 (1
991); including but not limited to the assay described in Schaub, Prostaglandins 35: 467-474, 1988.

Receptor / Ligand Activity The protein of the present invention may also exhibit activity as a receptor, receptor ligand or inhibitor or agonist of receptor / ligand interaction. Examples of such receptors and ligands include cytokine receptors and their ligands, receptor kinases and their ligands, receptor phosphatases and their ligands, cells-
Receptors involved in cell interactions and their ligands (such as cell adhesion molecules (selectins, integrins and their ligands)) and receptors / ligands involved in antigen presentation, antigen recognition, development of cellular and humoral immune responses But not limited thereto. Receptors and ligands are also useful for screening potential peptide or small molecule inhibitors for related receptor / ligand interactions. The proteins of the present invention, including but not limited to receptor and ligand fragments, may themselves be useful as inhibitors of receptor / ligand interaction. The activity of the protein of the invention may be measured, inter alia, by the following method: Suitable assays for receptor-ligand activity are described in Current Protocols in Immunology, Ed by JE Coligan, AM Kruisbeek, D.
H. Margulies, EM Shevach, W. Strober, Pub. Greene Publishing Associ
ates and Wiley-Interscience (Chapter 7.28, Measurement of Cellular Adhes
ion under static conditions 7.28.1-7.28.22), Takai et al., Proc. Natl. A
cad. 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; Stolte
nborg et al., J. Immunol.Methods 175: 59-68, 1994; Stitt et al., Cell 8
0: 661-670, 1995, including, but not limited to.

Anti-inflammatory activity The protein of the present invention can exhibit anti-inflammatory activity. Anti-inflammatory activity involves stimulating cells that are involved in the stimulus response, resulting in cell-cell interactions (eg, attachment).
By inhibiting or promoting, thereby inhibiting or promoting the chemotaxis of cells involved in the inflammatory process, thereby inhibiting or promoting cell extravasation,
Alternatively, it is exerted by stimulating or suppressing the production of other factors that more directly inhibit or promote the inflammatory response. Symptoms of inflammation (
Includes chronic or acute symptoms, infection-related inflammation (such as septic shock, sepsis or systemic inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, lethal endotoxin injury, arthritis, complement mediated Super-acute rejection,
Nephritis, lung injury induced by cytokines or chemokines, inflammatory bowel disease, Crohn's disease or diseases resulting from overproduction of cytokines such as TNF or IL-1). . The proteins of the invention may also be useful for treating anaphylaxis and hypersensitivity to antigenic substances or materials.

Cadherin / Tumor Inhibition Inhibitory Activity Cadherin is a calcium-dependent adhesion molecule and appears to play a major role during development, particularly in determining specific cell types. Loss or alteration of normal cadherin expression can induce changes to cell adhesion properties associated with tumor growth and metastasis. Cadherin dysfunction has also been implicated in other human diseases, such as pemphigus vulgaris and pemphigus foliaceus (autoimmune cutaneous bullous disease), Crohn's disease, and some developmental abnormalities. The cadherin superfamily contains over 40 members, each with a different expression pattern. All members of the superfamily share common conserved extracellular repeats (cadherin domains), but show structural differences in other parts of the molecule. Calcium is necessary for their attachment, since the cadherin domains bind to calcium to form their tertiary structure. Since the small number of amino acids in the first cadherin domain provides the basis for homophilic attachment, modification of this recognition site can alter the specificity of cadherin, thereby recognizing only itself Instead, the mutant molecule will be able to bind to different cadherins. In addition, some cadherins are involved in heterophilic attachment with other cadherins. E-cadherin, a member of the cadherin superfamily, is expressed in epithelial cell types. Pathologically, if E-cadherin expression is lost in the tumor, the malignant cells become invasive and the cancer metastasizes. Transfection of a polynucleotide expressing E-cadhedrin into a cancer cell line can reverse the altered cell morphology, restore cell-to-cell and adherence to other substrates, reduce cell growth rates, By dramatically reducing the growth of dependent cells,
The cancer-related changes were reversed. Thus, reintroducing E-cadhedrin expression returns the carcinoma to a less advanced stage. Other cadherins may have the same inhibitory role in carcinomas from other tissue types. Therefore, cancer can be treated using the protein of the present invention having cadherin activity and the polynucleotide of the present invention encoding such a protein. Introducing such proteins or polynucleotides into cancer cells may reduce or eliminate the cancerous changes observed in these cells by providing normal cadherin expression. Cancer cells are also known to express cadherins of different tissue types than originally, so these cancer cells can invade different tissues and metastasize. The protein of the present invention having cadherin activity, and the polynucleotide of the present invention encoding such a protein replaces cadherin inappropriately expressed in these cells, restores normal cell adhesion properties, and reduces the tendency of cells to metastasize. Or can be removed. Furthermore, an antibody that recognizes and binds to cadherin can be obtained using the protein of the present invention having cadherin activity and the polynucleotide of the present invention encoding such a protein. Such antibodies can also be used to block the attachment of inappropriately expressed tumor cells cadherin, so that the cells do not form tumors elsewhere. Such anti-cadhedrin antibodies can be used as markers for cancer grade, pathological type, and prognosis. That is, as the cancer progresses, the expression of cadherin decreases, and by using a cadherin-binding antibody, the decrease of cadherin expression can be detected. Using a fragment of a protein of the present invention having cadherin activity, preferably a decapeptide at the cadherin recognition site, and a polynucleotide of the present invention encoding such a protein fragment, to bind to cadherin to prevent cadherin binding that produces undesirable effects. Can also block cadherin function. Further, using a fragment of the protein of the present invention having cadherin activity, preferably a truncated soluble cadherin fragment known to be stable in the circulatory system of cancer patients, and a polynucleotide encoding such a protein fragment, Correct cell-cell attachment can also be disrupted. Assays for cadherin adhesion and entry inhibitory activity are described in Hortsch et al. J Bio
l Chem 270 (32): 18809-18817, 1995; Miyaki et al. Oncogene 11: 2547-2552
, 1995; Ozawa et al. Cell 63: 1033-1038, 1990.

Tumor Inhibitory Activity In addition to the activities described above for the immunological treatment or prevention of tumors, the proteins of the present invention may exhibit other antitumor activities. Certain proteins directly or indirectly affect tumor growth (eg,
(Via ADCC). Certain proteins act on tumor tissue or progenitor tissue to inhibit the formation of tissue necessary to support tumor growth (eg, by inhibiting angiogenesis) and to inhibit tumor growth. Tumor inhibitory activity may be exhibited by causing the production of a factor, agent or cell type, or by removing or inhibiting a factor, agent or cell type that promotes tumor growth.

Other Activities The proteins of the present invention may exhibit one or more of the following additional activities or effects: Infectious agents, including but not limited to bacteria, viruses, fungi, and other parasites. Killing; height, weight, hair color, eye color, skin or other tissue pigmentation, or the size or form of an organ or body part (eg, breast augmentation or vice versa)
Effects on body characteristics (suppression or promotion); effects on digestion of consumed fat, protein or carbohydrates; appetite, libido, stress, cognition (including cognitive disorders), depression (including depressive disorders) Effects on behavioral characteristics, including but not limited to, violent behavior; providing analgesic or other pain-relieving effects; promoting differentiation and proliferation of embryonic stem cells in non-hematopoietic lineages; hormones or endocrine Activity; for enzymes,
Correcting enzyme deficiency and treating related disorders; treating hyperproliferative disorders (eg, psoriasis); immunoglobulin-like activity (eg, the ability to bind antibodies or complement);
And the ability to act as an antigen in a vaccine composition to generate an immune response to such proteins or other substances or entities that cross-react with such proteins.

Administration and Dose The protein of the present invention (which may be from any source, including but not limited to recombinant and non-recombinant methods) is mixed with a pharmaceutically acceptable carrier. And may be used in pharmaceutical compositions. Such compositions may contain (in addition to proteins and carriers) diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The term "pharmaceutically acceptable"
A non-toxic substance that does not interfere with the effectiveness of the biological activity of the active ingredient. The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition of the present invention comprises M-CSF, GM-CSF, T
NF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-
7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13,
IL-14, IL-15, IFN, TNF0, TNF1, TNF2, G-CSF
, Meg-CSF, thrombopoietin, stem cell factor, and cytokines such as erythropoietin, lymphokines, or other hematopoietic factors. The pharmaceutical composition may further contain other agents that enhance protein activity or supplement its activity or utility in therapy. Such additional factors and / or agents may be included in a pharmaceutical composition to exert a synergistic effect with the protein of the invention or to minimize side effects. Conversely, specific cytokines, lymphokines, other hematopoietic factors, thrombolytic factors or antithrombotic factors, or by incorporating the protein of the present invention into the formulation of anti-inflammatory agents, cytokines, lymphokines, other hematopoietic factors, thrombolytic factors or Side effects of antithrombotic factors or anti-inflammatory agents may be minimized. A protein of the invention may be active in multimers (eg, heterodimers or homodimers) or in complexes with itself or other proteins. Consequently, the pharmaceutical composition of the present invention may contain such a multimeric or complexed form of the protein of the present invention. The pharmaceutical composition of the present invention may be in the form of a complex of the protein of the present invention and a protein or peptide antigen. Protein and / or peptide antigens can be
It will deliver a stimulus signal to the lymphocytes. B lymphocytes will respond to antigen through their surface immunoglobulin receptor. T lymphocytes will respond to antigen through the T cell receptor (TCR) after presentation of the antigen by MHC proteins.
MHC and structurally related proteins, including those encoded by the host cell's class I and class II MHC genes, will serve to present peptide antigens to T lymphocytes. The antigen component is supplied as a purified MHC-peptide complex alone or with a costimulatory molecule that can directly signal T cells. Alternatively, antibodies capable of binding to surface immunoglobulins and other molecules on B cells can be mixed with the pharmaceutical compositions of the present invention. The pharmaceutical composition of the present invention may be in the form of a liposome, in which the protein of the present invention further comprises another pharmaceutically acceptable carrier, an amphipathic agent such as a lipid (in the form of an aggregate such as micelles in an aqueous solution). , Present as an insoluble monolayer, liquid crystal or lamellar layer). Suitable lipids for liposome formulations include, but are not limited to, monoglycerides, diglycerides, sulfatide, lysolecithin, phospholipids, saponins, bile acids, and the like. The preparation of such liposome formulations is within the level of ordinary skill in the art and is described, for example, in US Pat.
And 4737323, all of which are herein incorporated by reference. As used herein, the term "therapeutically effective amount" refers to the amount of each active ingredient of a pharmaceutical composition or method that is sufficient to show significant patient benefit, e.g., amelioration of symptoms of such symptoms, healing, increased rate of healing. Means the total amount. When used for an individual active ingredient administered alone, the term refers only to that ingredient. When used in a combination of active ingredients, it refers to the total amount of active ingredients that produces a therapeutic effect regardless of sequential or simultaneous administration. In practicing the therapeutic methods of the invention, a therapeutically effective amount of a protein of the invention is administered to a mammal having the condition to be treated. According to the method of the invention, the protein of the invention may be administered alone or with other therapeutic agents, such as cytokines, lymphokines or other hematopoietic factors. When co-administered with one or more cytokines, lymphokines or other hematopoietic factors, the proteins of the invention may be administered simultaneously or sequentially with cytokines, lymphokines, other hematopoietic factors, thrombolytic or antithrombotic factors. If administered sequentially, the attending physician will determine the appropriate order of administration for combinations of the proteins of the present invention with cytokines, lymphokines, other hematopoietic, thrombolytic or antithrombotic factors. The administration of the protein of the present invention in the pharmaceutical composition of the present invention or used in the practice of the present invention can be performed by various conventional methods, for example, oral feeding, inhalation, or intradermal, subcutaneous, or intravenous injection. Intravenous injection into a patient is preferred. When a therapeutically effective amount of a protein of the invention is administered orally, the protein of the 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 further contain a solid carrier such as gelatin or an adjuvant. Tablets, capsules, and powders contain about 5 to 95% of a protein of the present invention;
It preferably contains about 25-90% of the protein of the invention. When administered in liquid form, water, petroleum, oils of animal or vegetable origin, such as peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oils may be added. Liquid form pharmaceutical compositions may further contain physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. When administered in liquid form, the pharmaceutical composition contains about 0.5 to 90% by weight of the protein of the present invention, preferably about 1 to 50% of the protein of the present invention. When administering a therapeutically effective amount of a protein of the invention intravenously, intradermally or subcutaneously,
The protein of the present invention may be in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such a parenterally acceptable protein solution having appropriate pH, isotonicity, and stability is within the skill of the art. Preferred pharmaceutical compositions for intravenous, intradermal, or subcutaneous injection include, in addition to the protein of the present invention, sodium chloride for injection, Ringer's solution, dextrose for injection, dextrose and sodium chloride for injection, and Ringer's solution containing lactic acid for injection, Or it should contain other carriers known in the art. The pharmaceutical composition of the present invention may contain stabilizers, preservatives, buffers, antioxidants, or other additives known to those skilled in the art. The amount of the protein of the invention in the pharmaceutical composition of the invention depends on the nature and severity of the condition to be treated,
As well as the nature of the treatment the patient has already received. Ultimately, your doctor
The amount of the protein of the invention to be treated for each patient will be determined. First, the attending physician administers a low dose of the protein of the invention and observes the patient's response. Higher doses of the protein of the invention may be administered until the optimal therapeutic effect is obtained, with no further increase in the dose at the time the optimal therapeutic effect is obtained. Various pharmaceutical compositions for carrying out the method of the present invention contain about 0.01 μg to about 100 mg of the protein of the present invention (preferably,
It should contain from about 0.1 μg to about 10 mg per kg body weight, more preferably from 0.1 μg to about 1 mg per kg body weight. The duration of treatment by intravenous administration using the compositions of the present invention will vary with the severity of the disease to be treated and the condition and response of each patient. It is believed that the duration of each application of the protein of the invention ranges from 12 to 24 hours for continuous intravenous administration. Ultimately, the attending physician will decide the duration of intravenous treatment using the pharmaceutical composition of the present invention.

An animal may be immunized using the protein of the present invention to obtain polyclonal and monoclonal antibodies that specifically react with the protein of the present invention. Such antibodies may be obtained using whole proteins or fragments thereof as immunogens. The peptide immunogen may further include a cysteine residue at the carboxy terminus and binds to haptens such as keyhole limpet hemocyanin (KLH). Methods for synthesizing such peptides are known in the art, for example, RP Merrifield, J. Amer. Che.
m. Soc. 85, 2149-2154 (1963); JL Krstenansky, et. al., FEBS Lett. 211
, 10 (1987). A monoclonal antibody that binds to the protein of the present invention may be a useful diagnostic agent for immunodetection of the protein of the present invention. Neutralizing antibodies that bind to the protein of the present invention can be useful as therapeutics for conditions related to the protein of the present invention,
Further, it may be useful in the treatment of certain tumors in the treatment of some forms of cancer in which aberrant expression of the protein of the invention is involved. For cancer cells or white blood cells,
Neutralizing monoclonal antibodies against the protein of the present invention may be useful for detecting and preventing metastatic spread of cancer cells mediated by the protein of the present invention. For compositions of the invention useful for regenerating bone, cartilage, tendons or ligaments, the method of treatment comprises:
It involves administering the composition locally, systemically, or locally, such as an implant or device. When administered, the therapeutic composition used in the present invention is, of course, in a pyrogen-free, physiologically acceptable form. In addition, if desired, the composition may be encapsulated or injected in a viscous form and delivered to the site of bone, cartilage or tissue damage. Topical administration is suitable for wound healing and tissue repair. Therapeutically useful agents other than the protein of the present invention, which may be contained in the above composition, may be alternatively or additionally administered simultaneously or sequentially with the composition in the method of the present invention. Preferably, for bone and / or cartilage formation, the composition delivers the protein-containing composition to the site of bone and / or cartilage damage, provides a structure for bone and cartilage development, and more optimally. Will contain a matrix that can be absorbed into the body. Such matrices may be made of materials currently used for other implanted medical devices. The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The appropriate formulation will be determined by the particular application of the composition. Possible matrices for the composition may be biodegradable and chemically known calcium sulfate, tricalcium phosphate, hydroxyapatite, polylactic acid, polyglycolic acid and polyanhydride. Other usable materials are biodegradable and biologically well known, such as bone or skin collagen. Further, the matrix may contain pure proteins or extracellular matrix components.
Other possible matrices are not biodegradable, such as sintered hydroxyapatite, bioglass, aluminate, or other ceramics, but are chemically known. The matrix may comprise a combination of materials of the above type, for example polylactic acid and hydroxyapatite or collagen and tricalcium phosphate. The bioceramics may be modified in the composition, for example in calcium-aluminate-phosphate, and processed to pore size, particle size,
The particle shape and biodegradability may be varied. A 50:50 (molar weight) copolymer of lactic acid and glycolic acid in the form of porous particles having a diameter in the range of 150 to 800 microns is presently preferred. In some applications, it may be useful to use a sequestering agent, such as carboxymethylcellulose or an autologous clot, to prevent dissociation of the protein composition from the matrix. A preferred family of sequestering agents is cellulosic materials such as alkylcellulose (including hydroxyalkylcellulose), including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose; Most preferred are the cationic salts of (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly (ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymers and poly (vinyl alcohol). The amount of sequestering agent useful in the present invention is from 0.5 to 20% by weight, preferably from 1 to 10% by weight, based on the total formulation weight, and this amount prevents desorption of proteins from the polymer matrix. An amount that allows for proper handling of the composition, but not too much so that the progenitor cells infiltrate the matrix, thereby conferring the protein with the opportunity to promote the osteogenic activity of the progenitor cells. Not to be. In a further composition, the protein of the present invention may be mixed with other agents that are beneficial in treating bone and / or cartilage defects, wounds, or tissues. These agents include epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factors (TGF-α and TGF-β), and insulin-like growth factor (IGF). At present, the therapeutic compositions are also valuable for veterinary use. For details,
In addition to humans, livestock and thoroughbred horses are desirable patients for such treatments using the proteins of the present invention. The rules of administration of the protein-containing pharmaceutical composition used for tissue regeneration may include various factors that alter the action of the protein, such as the tissue weight desired to be formed, the site of damage, the condition of the damaged tissue, the size of the wound, The physician will decide on the type of tissue required (eg, bone), patient age, gender, and diet, severity of infection, duration of administration, and other clinical factors. Dosages may vary depending on the type of matrix used for reconstitution and the inclusion of other proteins in the pharmaceutical composition. For example, IGF
The addition of other known growth factors, such as I (insulin-like growth factor I), to the final composition can also affect the dose. Progress can be monitored by periodically assessing tissue / bone growth and / or repair, for example, by X-ray, histomorphological examination and tetracycline labeling. The polynucleotide of the present invention can also be used for gene therapy. Such polynucleotides can be introduced into cells in vivo or ex vivo and expressed in a mammalian subject. The polynucleotides of the present invention may be administered by other known methods for introducing nucleic acids into cells or organisms, including but not limited to viral vectors or in the form of naked DNA. ). The cells may be cultured and grown ex vivo in the presence of the protein of the present invention, or may have a desired effect on such cells, or may have a desired activity in such cells. The treated cells can then be introduced in vivo for therapeutic purposes. The patents and publications cited herein are considered to be entirely incorporated herein by reference.

[0056]

[Sequence list]

[Brief description of the drawings]

FIG. 1A is a schematic representation of the pED6 vector used for depositing the clones disclosed herein.

FIG. 1B shows the pNOTs used to deposit the clones disclosed herein.
1 schematically shows a vector.

──────────────────────────────────────────────────続 き Continuation of front page (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE ), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE, GH, GM, HR, HU, ID, IL, IS, JP, KE, KG, KP , KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, UZ, VN, YU, ZW (72) Inventor John M. McCoy United States 01867 Massachusetts Reading, Howard Street 56 (72) Inventor Edward Earle Lavary United States 01451 Ann Lee Road, Harvard, Mass. 113 (72) Inventor Lisa A. Collins-Lacy United States 01720 Acton, School Street 124, Massachusetts, United States 01720 Cheryl Inventor Evans United States 01915 Brighton Drive 307, Beverly, MA (72) Inventor Davi De Marberg, United States 01720 Massachusetts, Acton, Orchard Drive No. 2 (72) Inventor Maurice Tracy United States 02167 Massachusetts Chestnut Hill, Walcott Rhode No. 93 (72) Inventor Vicky Spalding United States 01821 Massachusetts Billerica, Meadowbank Road No.11 F term (reference) 4B024 AA01 AA11 BA80 CA01 CA04 DA02 EA04 FA17 GA11 HA01 HA03 4B064 AG01 CA10 CA19 CC24 DA01 DA13 4B065 AA90Y AB01 BA02 CA24 CA43 CA46 4H045 AA10 BA10 CA40 FA72 EA50

Claims (38)

[Claims] 1. A polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1; (b) a polynucleotide comprising the nucleotide sequence from nucleotide 54 to nucleotide 1283 of SEQ ID NO: 1; A polynucleotide comprising the nucleotide sequence from nucleotide 648 to nucleotide 1283; (d) a polynucleotide comprising the nucleotide sequence from nucleotide 458 to nucleotide 947 of SEQ ID NO: 1; (e) clone AY421 2 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of (a) clone AY4212 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of (c) clone AY4212 deposited under accession number ATCC 98145.
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of (h) clone AY4212, deposited under accession number ATCC 98145.
A polynucleotide encoding a mature protein encoded by the cDNA insert of (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 2; (j) a biologically active SEQ ID NO: A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of (2); (k) a polynucleotide which is an allelic variant of the polynucleotide of (a) to (h); A) a polynucleotide that encodes a species homolog of the protein; and (m) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides specified in (a) to (j). A composition comprising an isolated polynucleotide selected from the group consisting of: 2. The composition of claim 1, wherein said polynucleotide is operably linked to at least one expression control sequence. 3. A host cell transformed with the composition of claim 2. 4. The host cell according to claim 3, wherein said cell is a mammalian cell. 5. The encoded by the composition of claim 2, comprising: (a) growing the culture of the host cell of claim 3 in a suitable medium; and (b) purifying the protein from the culture. Method for producing protein. 6. A protein produced by the method of claim 5. 7. An isolated polynucleotide encoding the protein of claim 6. 8. A clone AY deposited under accession number ATCC 98145.
The polynucleotide of claim 7, comprising the 4212 cDNA insert. 9. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a) a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 2 (B) an amino acid sequence from amino acid 180 to amino acid 298 of SEQ ID NO: 2; (c) a fragment of the amino acid sequence of SEQ ID NO: 2; and (d) clone AY421 2 deposited under accession number ATCC 98145.
Amino acid sequence encoded by the cDNA insert of FIG. 10. The composition of claim 9, wherein said protein comprises the amino acid sequence of SEQ ID NO: 2. 11. The protein of claim 1, wherein the protein is amino acid 180 to amino acid 29 of SEQ ID NO: 2.
10. The composition of claim 9, comprising up to 8 amino acid sequences. 12. The composition of claim 9, further comprising a pharmaceutically acceptable carrier. 13. A method of preventing, treating or ameliorating a medical condition comprising administering to a mammalian subject a therapeutically effective amount of the composition of claim 12. 14. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 1. 15. A polynucleotide comprising the nucleotide sequence of SEQ ID NO: 3; (b) a polynucleotide comprising nucleotides 720 to 974 of SEQ ID NO: 3; (c) a polynucleotide comprising nucleotides 715 of SEQ ID NO: 3 A polynucleotide comprising up to nucleotide 947; (d) clone BV2782 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of (c) BV2782 deposited under accession number ATCC 98145
A polynucleotide encoding the full length protein encoded by the cDNA insert of (b) clone BV2782 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of: (g) clone BV2782 deposited under accession number ATCC 98145.
(H) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 4; (i) an amino acid sequence of SEQ ID NO: 4 having biological activity (J) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; (k) a species homology 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) to (i). A composition comprising: 16. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 4 (B) an amino acid sequence from amino acid 1 to amino acid 76 of SEQ ID NO: 4; (c) a fragment of the amino acid sequence of SEQ ID NO: 4; and (d) clone BV2782 deposited under accession number ATCC 98145.
Amino acid sequence encoded by the cDNA insert of FIG. 17. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 3. 18. A polynucleotide comprising: (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 5; (b) a polynucleotide comprising the nucleotide sequence from nucleotide 36 to nucleotide 968 of SEQ ID NO: 5; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 5 A polynucleotide comprising the nucleotide sequence of from nucleotide 340 to nucleotide 717; (d) a polynucleotide comprising the nucleotide sequence of the full-length protein coding sequence of clone C5441 deposited under accession number ATCC98145; (e) a polynucleotide comprising accession number ATCC98145 A polynucleotide encoding the full-length protein encoded by the cDNA insert of clone C5441; (f) a sequence encoding the mature protein coding sequence of clone C5441 deposited under accession number ATCC 98145; (G) a polynucleotide encoding the mature protein encoded by the cDNA insert of clone C5441 deposited under accession number ATCC 98145; (h) a protein comprising the amino acid sequence of SEQ ID NO: 6 A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 6 having biological activity; and (j) a polynucleotide encoding the above (a) to (g). A polynucleotide that is an allelic variant of a nucleotide; (k) a polynucleotide that encodes a species homolog of the protein of (h) or (i) above; and (1) under stringent conditions (a)-(i). A) a polynucleotide that is capable of hybridizing to any one of the polynucleotides specified in) A composition comprising an isolated polynucleotide selected from the group consisting of: 19. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a) a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 6 (B) an amino acid sequence from amino acid 103 to amino acid 227 of SEQ ID NO: 6; (c) a fragment of the amino acid sequence of SEQ ID NO: 6; and (d) a cDNA insert of clone C5441 deposited under accession number ATCC 98145. The amino acid sequence encoded by 20. An isolated gene corresponding to SEQ ID NO: 5 cDNA sequence. 21. (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 7; (b) a polynucleotide comprising the nucleotide sequence from nucleotide 151 to nucleotide 1398 of SEQ ID NO: 7; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 7 A polynucleotide comprising the nucleotide sequence from nucleotide 637 to nucleotide 1398; (d) a polynucleotide comprising the nucleotide sequence from nucleotide 255 to nucleotide 429 of SEQ ID NO: 7; (e) clone CC332 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of SEQ ID NO: 3; (f) clone CC332 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of No. 3; (g) clone CC332 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of 3; (h) clone CC332 3 deposited under accession number ATCC 98145
A polynucleotide encoding a mature protein encoded by the cDNA insert of SEQ ID NO: 3; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 8; (j) a SEQ ID NO having biological activity : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of (8); (k) a polynucleotide which is an allelic variant of the polynucleotide of (a) to (h); (l) a (i) or ( j) a polynucleotide encoding a species homolog of the protein; and (m) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a) to (j). A composition comprising an isolated polynucleotide selected from the group. 22. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: (a) an amino acid of SEQ ID NO: 8; (B) an amino acid sequence from amino acid 36 to amino acid 93 of SEQ ID NO: 8; (c) a fragment of the amino acid sequence of SEQ ID NO: 8; and (d) clone CC332_3 deposited under accession number ATCC 98145.
Amino acid sequence encoded by the cDNA insert of No. 3. 23. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 7. 24. (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 32; (b) a polynucleotide comprising the nucleotide sequence from nucleotide 21 to nucleotide 410 of SEQ ID NO: 32; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 32 A polynucleotide comprising the nucleotide sequence from nucleotide 144 to nucleotide 410; (d) a polynucleotide comprising the nucleotide sequence from nucleotide 68 to nucleotide 368 of SEQ ID NO: 32; (e) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of SEQ ID NO: 0; (f) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide encoding the full-length protein encoded by the cDNA insert of No. 0; (g) clone CC365 4 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of SEQ ID NO: 0; (h) clone CC365_4 deposited under accession number ATCC 98145
A polynucleotide encoding a mature protein encoded by a cDNA insert of 0; (i) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 33; (j) a SEQ ID NO having biological activity : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of SEQ ID NO: 33
(K) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (h) above; (l) a species homology 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). A composition comprising a polynucleotide. 25. A method for producing an isolated polynucleotide, comprising:
a) a method selected from the group consisting of: (b) (a) a method comprising the following steps: (i) a 6X SSC at 65 ° C. in a nucleotide sequence selected from the group consisting of: Preparing one or more polynucleotide probes that hybridize; (aa) SEQ ID NO: 32 (excluding the poly (A) tail at the 3 ′ end of SEQ ID NO: 32); and (ab) deposited as ATCC 98145. Clone CC365 40
(Ii) hybridizing the probe (s) to human DNA; (iii) isolating the DNA polynucleotide detected with the probe (s); and (B) a method comprising the steps of: (i) preparing one or more polynucleotide primers that hybridize in 6X SSC at 65 ° C. to a nucleotide sequence selected from the group consisting of: (Ba) SEQ ID NO: 32 (excluding the poly (A) tail at the 3 ′ end of SEQ ID NO: 32); and (bb) Clone CC36540 deposited as ATCC 98145
(Ii) hybridizing the primer (s) to human DNA; (iii) amplifying the human DNA sequence; then (iv) the polynucleotide of step (b) (iii) Isolating the product. 26. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a) a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 33; (B) an amino acid sequence from amino acid 17 to amino acid 116 of SEQ ID NO: 33; (c) a fragment of the amino acid sequence of SEQ ID NO: 33 comprising eight consecutive amino acids of SEQ ID NO: 33; and (d) ) Clone CC3654 deposited under accession number ATCC 98145
Amino acid sequence encoded by 0 cDNA insert. 27. (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 12; (b) a polynucleotide comprising the nucleotide sequence of nucleotide 769 to nucleotide 966 of SEQ ID NO: 12; (c) deposited as accession number ATCC 98145 A polynucleotide comprising the nucleotide sequence of the cloned full-length protein coding sequence of clone CG684; (d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone CG684 deposited under accession number ATCC 98145; A) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone CG684 deposited under accession number ATCC 98145; (f) clone CG6 deposited under accession number ATCC 98145 A polynucleotide encoding a mature protein encoded by the cDNA insert of No. 4; (g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 13; (h) a SEQ ID NO having biological activity : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of 13; (i) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (f) above; (j) a (g) or (g) h) a polynucleotide encoding a species homolog of the protein; and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a) to (h). A composition comprising an isolated polynucleotide selected from the group. 28. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 12. 29. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of SEQ ID NO: 13; (B) an amino acid sequence from amino acid 18 to amino acid 57 of SEQ ID NO: 13; (c) a fragment of the amino acid sequence of SEQ ID NO: 13; and (d) a cDNA insert of clone CG684 deposited under accession number ATCC 98145. The amino acid sequence encoded by 30. (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 14; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 14 from nucleotide 1042 to nucleotide 1389; (c) a deposit comprising accession number ATCC 98145. A polynucleotide comprising the nucleotide sequence of the cloned full length protein coding sequence of clone D3291; (d) a polynucleotide encoding the full length protein encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145; A) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone D3291 deposited under accession number ATCC 98145; (f) clone D deposited under accession number ATCC 98145 A polynucleotide encoding a mature protein encoded by the 291 cDNA insert; (g) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 15; (h) a sequence having biological activity A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of No. 15; (i) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (f); (j) a (g) or (H) a polynucleotide encoding a species homolog of the protein of (h); and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides (a) to (h). A composition comprising an isolated polynucleotide selected from the group consisting of: 31. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 14. 32. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 15 (B) a fragment of the amino acid sequence of SEQ ID NO: 15; and (c) the amino acid sequence encoded by the cDNA insert of clone D3291 deposited under accession number ATCC 98145. 33. a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 18; (b) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 18 from nucleotide 279 to nucleotide 515; (c) a deposit comprising accession number ATCC 98145. A polynucleotide comprising the nucleotide sequence of the cloned full-length protein coding sequence of clone H6983; (d) a polynucleotide encoding the full-length protein encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145; A) a polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of clone H6983 deposited under accession number ATCC 98145; (f) clone H69 deposited under accession number ATCC 98145 (G) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 19; (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 19; : A polynucleotide encoding a protein comprising a fragment of the amino acid sequence of 19; (i) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (f) above; (j) a (g) or (g) h) a polynucleotide encoding a species homolog of the protein; and (k) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a) to (h). A composition comprising an isolated polynucleotide selected from the group. 34. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 17 or SEQ ID NO: 18. 35. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a protein substantially free of other mammalian proteins: (a) an amino acid of SEQ ID NO: 19 (B) a fragment of the amino acid sequence of SEQ ID NO: 19; and (c) the amino acid sequence encoded by the cDNA insert of clone H6983 deposited under accession number ATCC 98145. 36. (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 21; (b) a polynucleotide comprising the nucleotide sequence from nucleotide 199 to nucleotide 1155 of SEQ ID NO: 21; (c) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 21 A polynucleotide comprising the nucleotide sequence from nucleotide 304 to nucleotide 1155; (d) clone H96320 deposited under accession number ATCC 98145
A polynucleotide comprising the nucleotide sequence of the full length protein coding sequence of (c) H96320 deposited under accession number ATCC 98145.
A polynucleotide encoding the full-length protein encoded by the cDNA insert of (h) clone H96320 deposited under accession number ATCC 98145;
A polynucleotide comprising the nucleotide sequence of the mature protein coding sequence of (c) H96320 deposited under accession number ATCC 98145.
A polynucleotide encoding a mature protein encoded by the cDNA insert of (h) a polynucleotide encoding a protein comprising the amino acid sequence of SEQ ID NO: 22; (i) a SEQ ID NO: having biological activity A polynucleotide encoding a protein comprising a fragment of 22 amino acid sequences; (j) a polynucleotide that is an allelic variant of the polynucleotide of (a) to (g) above; (k) a (h) or (i) A) a polynucleotide that encodes a species homolog of the protein of (a); and (1) a polynucleotide that hybridizes under stringent conditions to any one of the polynucleotides of (a) to (i). A composition comprising an isolated polynucleotide selected from the group consisting of: 37. An isolated gene corresponding to the cDNA sequence of SEQ ID NO: 21. 38. A composition comprising a protein comprising an amino acid sequence selected from the group consisting of: a) a protein substantially free of other mammalian proteins: (a) the amino acid of SEQ ID NO: 22 (B) an amino acid sequence from amino acid 19 to amino acid 84 of SEQ ID NO: 22; (c) a fragment of the amino acid sequence of SEQ ID NO: 22; and (d) clone H9620 deposited under accession number ATCC 98145.
Amino acid sequence encoded by the cDNA insert of FIG.