Mammalian cytokines; receptors; related reagents and methods

• US 20050014199A1
• Filed: 08/25/2004
• Published: 01/20/2005
• Est. Priority Date: 11/10/2000
• Status: Active Grant

• Current Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Original Assignee: Schering Corporation (Merck & Co., Inc.)
• Inventors: de Waal Malefyt, Rene, Liu, Yong-Jun, Bazan, J. Fernando, Kastelein, Robert A., Reche-Gallardo, Pedro A., Soumelis, Vassilli
• Granted Patent: US 7,071,308 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/7155 for interleukins [IL]
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: G01N 2333/54 Interleukins [IL]
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/6869 Interleukin

Abstract

Nucleic acids encoding mammalian cytokine receptor, e.g., for cytokine IL-B50, purified proteins and fragments thereof Antibodies, both polyclonal and monoclonal, are also provided. Methods of using the compositions for both diagnostic and therapeutic utilities are described.

Claims

1. A method of producing a ligand:

receptor complex, comprising contacting;

a) a substantially pure or recombinant mammalian IL-B50 with a receptor comprising the IL-7Rα

or the R52 subunit;

b) a mammalian IL-B50 with a receptor comprising a substantially pure or recombinant IL-7Rα

subunit;

or

c) a mammalian IL-B50 with a receptor comprising a substantially pure or recombinant Rδ

2 subunit;

thereby allowing said complex to form.

2. The method of claim 1, wherein:

a) said mammalian IL-B50 is primate IL-B50;

b) said complex formation results in signal transduction, STAT activation, or TARC, PARC or MDC expression;

c) said receptor is on a cell;

d) said receptor comprises both IL-7Rα and

Rδ

2 subunit;

e) said complex formation results in a physiological change in the cell expressing said receptor;

f) said contacting is in combination with a proliferative agent, cytokine, or chemokine;

or

g) said contacting allows quantitative detection of said ligand.

3. The method of claim 2, wherein said receptor is on a hematopoietic cell, including a lymphoid lineage cell, myeloid, or dendritic cell.

4. A method of modulating physiology or development of an IL-7Rα

or Rδ

2 expressing cell comprising contacting said cell to an exogenous agonist or antagonist of a mammalian IL-B50.

5. The method of claim 4, wherein:

A) said antagonist is;

1) an antibody which neutralizes said mammalian IL-B50;

2) amutein of said IL-B50;

or

3) an antibody which binds to IL-7Rα

or Rδ

2 or a complex comprising both;

B) said physiology is selected from;

1) proliferation;

2) lymphoid or dendritic lineage cell development;

3) antigen presentation;

or

4) production of inflammatory mediators, including cytokines, chemokines, or adhesion molecules;

or

C) said cell is a hematopoietic cell.

6. The method of claim 4, wherein:

a) said antagonist is an antibody and said physiology is hematopoietic cell proliferation;

b) said agonist is IL-B50 and said physiology is hematopoietic cell differentiation;

or

c) said physiology is antigen presentation.

7. The method of claim 4, wherein said modulating is blocking, and said physiology is lymphoid lineage cell proliferation.

8. A method of modulating a signal to a cell mediated by IL-B50 comprising contacting said cell to an administered agonist or antagonist of IL-B50.

9. The method of claim 8, wherein said modulating is inhibiting, and said signal is a proliferation signal.

10. The method of claim 9, wherein:

a) said antagonist is a neutralizing antibody to IL-7Rα

or the Rδ

2 subunit or a complex comprising said subunits;

b) said agonist or antagonist is administered in combination with another antagonist or agonist of IL-B50;

or

c) said agonist or antagonist is administered in combination with a growth factor, cytokine, chemokine, or immune adjuvant.

11. The method of claim 9, wherein said contacting is with another anti-proliferative agent or treatment.

12. A method of selectively labeling a population of cells, said method comprising contacting said cells with an antibody which binds:

a) IL-7Rα

;

b) Rδ

2;

or

c) a complex comprising one of said subunits;

thereby resulting in the identification of cells expressing said subunit or complex.

13. The method of claim 12, wherein:

a) said contacting results in modulation of STAT activation;

b) said labeling allows purification of IL-7Rα

or Rδ

2 subunit expressing cells;

or

c) said labeling allows depletion of IL-7Rα

or Rδ

2 subunit expressing cells.

14. A population of cells made by the method of claim 13.

15. The population of claim 14, which are prepared by Fluorescent Activated Cell Sorting.

16. A method of testing a compound for ability to affect receptor-ligand interaction, said method comprising comparing the interaction of a receptor complex comprising IL-7Rα

and/or Rδ

8 subunit with IL-B50 in the presence and absence of said compound.

17. The method of claim 16, wherein said compound is an antibody which binds one of

a) IL-7Rα

b) Rδ

2 subunit;

c) a receptor comprising IL-7R and/or Rδ

2;

or

d) IL-B50.

18. An isolated or recombinant protein complex comprising:

a) at least 15 contiguous amino acid residues of SEQ ID NO;

2 and at least 15 contiguous amino acid residues of SEQ ID NO;

4;

b) at least two distinct segments of at least 8 contiguous amino acid residues of SEQ ID NO;

2 and at least two distinct segments of at least 8 contiguous amino acid residues of SEQ ID NO;

4;

or

c) at least one segment at least 21 contiguous nucleotides of SEQ ID NO;

1 and at least one segment at least 21 contiguous nucleotides of SEQ ID NO;

3.

19. The complex of claim 18, wherein:

a) one of said segments of SEQ ID NO;

2 is from the extracellular portion of said sequence;

b) one of said segments of SEQ ID NO;

4 is from the extracellular portion of said sequence;

or

c) said polypeptide comprises the mature SEQ ID NO;

2 and the mature SEQ ID NO;

4 sequences.

20. An isolated or recombinant polynucleotide encoding said complex of claim 19, wherein:

a) said polynucleotide comprises a deoxyribonucleotide;

b) said polynucleotide comprises a ribonucleotide;

or

c) at least one of said segments is operably linked to a promoter.

21. A binding compound comprising an antigen binding portion from an antibody which binds with selectivity to a polypeptide comprising at least 12 contiguous amino acid residues of SEQ ID NO:

2 and at least 12 contiguous amino acid residues of SEQ ID NO;

4;

said polypeptide encoded by a nucleotide of claim 20.