Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Lymphokines – e.g. – interferons – interlukins – etc.
Reexamination Certificate
1999-01-15
2002-11-26
Eyler, Yvonne (Department: 1647)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Lymphokines, e.g., interferons, interlukins, etc.
C424S085100
Reexamination Certificate
active
06486301
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a novel human gene encoding a polypeptide which is a novel human cytokine. More specifically, isolated nucleic acid molecules are provided encoding a human polypeptide named Interleukin 20, hereinafter referred to as “IL-20”. IL-20 polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. Also provided are diagnostic methods for detecting disorders related to the immune system, and therapeutic methods for treating such disorders.
The invention further relates to screening methods for identifying agonists and antagonists of IL-20 activity.
BACKGROUND OF THE INVENTION
Cytokines typically exert their respective biochemical and physiological effects by binding to specific receptor molecules. Receptor binding will then stimulate specific signal transduction pathways (Kishimoto, T., et al.,
Cell
76:253-262 (1994). The specific interactions of cytokines with their receptors are often the primary regulators of a wide variety of cellular processes including activation, proliferation, and differentiation (Arai, K.-I, et al.,
Ann. Rev. Biochem
. 59:783-836 (1990); Paul, W. E. and Seder, R. A.,
Cell
76:241-251 (1994)).
Human interleukin (IL)-17 was only recently identified. IL-17 is a 155 amino acid polypetide which was molecularly cloned from a CD4+ T-cell cDNA library (Yao, Z., et al.,
J. Immunol
. 155:5483-5486 (1995)). The IL-17 polypeptide contains an N-terminal signal peptide and contains approximately 72% identity at the amino acid level with a T-cell trophic herpesvirus saimiri (HVS) gene designated HVS13. High levels of IL-17 are secreted from CD4-positive primary peripheral blood leukocytes (PBL) upon stimulation (Yao, Z., et al.,
Immunity
3:811-821 (1995)). Treatment of fibroblasts with IL-17, HVS13, or another murine homologue, designated CTLA8, activate signal transduction pathways and result in the stimulation of the NF-&kgr;B transcription factor family, the secretion of IL-6, and the costimulation of T-cell proliferation (Yao, Z., et al.,
Immunity
3:811-821 (1995)).
An HVS13-Fc fusion protein was used to isolate a murine IL-17 receptor molecule which does not appear to belong to any of the previously described cytokine receptor families (Yao, Z., et al.,
Immunity
3:811-821 (1995)). The murine IL-17 receptor (mIL-17R) is predicted to encode a type I transmembrane protein of 864 amino acids with an apparent molecular mass of 97.8 kDa. mIL-17R is predicted to possess an N-terminal signal peptide with a cleavage site between alanine-31 and serine-32. The molecule also contains a 291 amino acid extracellular domain, a 21 amino acid transmembrane domain, and a 521 amino acid cytoplasmic tail. A soluble recombinant IL-17R molecule consisting of 323 amino acids of the extracellular domain of IL-17R fused to the Fc portion of human immunoglobulin IgG1 was able to significantly inhibit IL-17-induced IL-6 production by murine NIH-3T3 cells (supra).
Interestingly, the expression of the IL-17 gene is highly restricted. It is typically observed primarily in activated T-lymphocyte memory cells (Broxmeyer, H.
J. Exp. Med
. 183:2411-2415 (1996); Fossiez, F., et al.,
J. Exp. Med
. 183:2593-2603 (1996)). Conversely, the IL-17 receptor appears to be expressed in a large number of cells and tissues (Rouvier, E., et al.,
J. Immunol
. 150:5445-5456 (1993); Yao, Z., et al.,
J. Immunol
. 155:5483-5486 (1995)). It remains to be seen, however, if IL-17 itself can play an autocrine role in the expression of IL-17. IL-17 has been implicated as a causitive agent in the expression of IL-6, IL-8, G-CSF, Prostaglandin E (PGE
2
), and intracellular adhesion molecule (ICAM)-1 (Fossiez, F., supra; Yao, Z., et al.,
Immunity
3:811-821 (1995)). Each of these molecules possesses highly relevent and potentially therapeutically valuable properties. For instance, IL-6 is involved in the regulation of hematopoietic stem and progenitor cell growth and expansion (Ikebuchi, K., et al.,
Proc. Natl. Acad. Sci. USA
84:9035-9039 (1987); Gentile, P. and Broxmeyer, H. E.
Ann. N.Y. Acad. Sci. USA
628:74-83 (1991)). IL-8 exhibits a myelosuppressive activity for stem cells and immature subsets of myeloid progenitors (Broxmeyer, H. E., et al.,
Ann. Hematol
. 71:235-246 (1995); Daly, T. J., et al.,
J. Biol. Chem
. 270:23282-23292 (1995)). G-CSF acts both early and late to activate and stimulate hematopoiesis in general, and more specifically on neutrophil hematopoiesis, while PGE
2
enhances erythropoiesis, suppresses lymphopoiesis and myelopoiesis in general, and strongly suppresses monocytopoiesis (Broxmeyer, H. E.
Amer. J. Ped. Hematol./Oncol
. 14:22-30 (1992); Broxmeyer, H. E. and Williams, D. E.
CRC Crit. Rev. Oncol./Hematol
. 8:173-226 (1988)).
Thus, there is a need for polypeptides that function as immunoregulatory molecules and, thereby, function in the transfer of an extracelluIlar signal ultimately to the nucleus of the cell, since disturbances of such regulation may be involved in disorders relating to cellular activation, hemostasis, angiogenesis, tumor metastasis, cellular migration and ovulation, as well as neurogenesis. Therefore, there is a need for identification and characterization of such human polypeptides which can play a role in detecting, preventing, ameliorating or correcting such disorders.
SUMMARY OF THE INVENTION
The present invention provides isolated nucleic acid molecules comprising a polynucleotide encoding at least a portion of the IL-20 polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 or the complete amino acid sequence encoded by the cDNA clone deposited as plasmid DNA as ATCC Deposit Number 209232 on Aug. 29, 1997. The nucleotide sequence determined by sequencing the deposited IL-20 clone, which is shown in
FIG. 1
(SEQ ID NO:1), contains an open reading frame encoding a complete polypeptide of 180 amino acid residues, including an initiation codon encoding an N-terminal methionine at nucleotide positions 45-47, and a predicted molecular weight of about 20.4 kDa. Nucleic acid molecules of the invention include those encoding the complete amino acid sequence excepting the N-terminal methionine shown in SEQ ID NO:2, or the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone in ATCC Deposit Number 209232, which molecules also can encode additional amino acids fused to the N-terminus of the IL-20 amino acid sequence.
The present invention also provides isolated nucleic acid molecules comprising a polynucleotide encoding at least a portion of the IL-20 polypeptide having the complete amino acid sequence shown in SEQ ID NO:15 or the complete amino acid sequence encoded by the cDNA clone deposited in a pool of 50 distinct plasmid DNA molecules as ATCC Deposit Number 209138 on Jul. 3, 1997. The sense and antisense nucleotide sequences determined by 209138 on Jul. 3, 1997. The sense and antisense nucleotide sequences determined by respectively, contain an open reading frame in the sense sequence (SEQ ID NO:28) encoding a polypeptide of 118 amino acid residues, including an initiation codon encoding an N-terminal methionine at nucleotide positions 59-61. Nucleic acid molecules of the invention include those encoding the complete amino acid sequence excepting the N-terminal methionine shown in SEQ ID NO:15, or the complete amino acid sequence excepting the N-terminal methionine encoded by the cDNA clone in ATCC Deposit Number 209138, which molecules also can encode additional amino acids fused to the N-terminus of the IL-20 amino acid sequence. Nucleic acid molecules of the invention further include those encoding any of the N-terminal and/or C-terminal IL-20 deletion mutations with termini of n
3
and/or m
3
as set forth below.
IL-20 has a predicted leader sequence of 20 amino acids underlined in
FIG. 1
; and the amino acid sequence of the predicted mature IL-20 protein is also shown in
FIG. 1
, as amino acid residues 21-180 and as residues 1-160 in SEQ ID NO:2. The encoded polypeptide also has a predict
Duan D. Roxanne
Ebner Reinhard
Florence Kimberly A.
Hu Jing-Shan
Murphy Marianne
Eyler Yvonne
Hamud Fozia
Human Genome Sciences Inc.
Human Genome Sciences Inc.
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