Claims
- 1. A soluble CTLA4 mutant molecule which binds CD80 and/or CD86 comprising an extracellular domain of CTLA4 so that (a) an alanine at position +29 is substituted with an amino acid selected from the group consisting of tyrosine, leucine, phenylalanine, tryptophan, and threonine, and (b) a leucine at position +104 is substituted with a glutamic acid.
- 2. The soluble CTLA4 mutant molecule of claim 1 further comprising an amino acid sequence which alters the solubility, affinity or valency of the soluble CTLA4 mutant molecule for binding to the CD80 and/or CD86 molecule.
- 3. The soluble CTLA4 mutant molecule of claim 2, wherein the amino acid sequence comprises a human immunoglobulin constant region.
- 4. The soluble CTLA4 mutant molecule of claim 2 further comprising an amino acid sequence which permits secretion of the soluble CTLA4 mutant molecule.
- 5. The soluble CTLA4 mutant molecule of claim 4, wherein the amino acid sequence comprises an oncostatin M signal peptide.
- 6. The soluble CTLA4 mutant molecule of claim 1 comprising methionine at position +1 through aspartic acid at position +124 as shown in FIG. 7.
- 7. The soluble CTLA4 mutant molecule of claim 1, comprising alanine at position −1 through aspartic acid at position +124 as shown in FIG. 7.
- 8. The soluble CTLA4 mutant molecule of claim 3, wherein the human immunoglobulin constant region is mutated to include a cysteine at position +130 substituted with a serine, a cysteine at position +136 substituted with a serine, a cysteine at position +139 substituted with a serine, and a proline at position +148 substituted with serine, as shown in FIG. 7.
- 9. A soluble CTLA4 mutant molecule which binds with higher avidity to CD80 and/or CD86 than CTLA4, comprising an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 10. A soluble CTLA4 mutant molecule which binds with higher avidity to the CD80 and/or CD86 than CTLA4, comprising an extracellular domain of CTLA4, wherein in the extracellular domain, leucine at position +104 is substituted with glutamic acid as shown in FIG. 8.
- 11. A nucleic acid molecule comprising a nucleotide sequence encoding the amino acid sequence corresponding to the soluble CTLA4 mutant molecule of claim 1.
- 12. The nucleic acid molecule of claim 11 having the sequence beginning with adenine at nucleotide position +1 and ending with adenine at +1071 as shown in FIG. 7 or 8.
- 13. The nucleic acid molecule of claim 11 having the sequence beginning with guanidine at −3 and ending at adenine at +1071 as shown in FIG. 7 or 8.
- 14. A vector comprising the nucleotide sequence of claim 11.
- 15. A host vector system comprising a vector of claim 14 in a suitable host cell.
- 16. The host vector system of claim 15, wherein the suitable host cell is a bacterial cell or a eukaryotic cell.
- 17. A method for producing a soluble CTLA mutant protein comprising growing the host vector system of claim 15 so as to produce the protein in the host cell and recovering the protein so produced.
- 18. A soluble CTLA mutant protein produced by the method of claim 17.
- 19. A method for regulating a T cell interaction with a CD80 and/or CD86 positive cell comprising contacting the CD80 and/or CD86 positive cell with the soluble CTLA4 mutant molecule of claim 1 so as to form a CTLA4 mutant molecule/CD80 or a CTLA4 mutant molecule/CD86 complex, the complex interfering with interaction between the T cell and the CD80 and/or CD86 positive cell.
- 20. The method of claim 20, wherein the soluble CTLA4 mutant comprises an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 21. The method of claim 20, wherein the soluble CTLA4 mutant molecule comprises an extracellular domain of CTLA4, wherein in the extracellular domain, leucine at position +104 is substituted with glutamic acid as shown in FIG. 8.
- 22. The method of claim 20, wherein the CD80 and/or CD86 positive cell is contacted with a fragment or a derivative of the soluble CTLA4 mutant molecule.
- 23. The method of claim 20, wherein the CD80 and/or CD86 positive cell is an APC cell.
- 24. The method of claim 20, wherein the interaction of the CTLA4-positive T cells with the CD80 and CD86 positive cells is inhibited.
- 25. A method for treating immune system diseases mediated by T cell interactions with CD80 and/or CD86 positive cells comprising administering to a subject the soluble CTLA4 mutant molecule of claim 1 to regulate T cell interactions with the CD86 positive cells.
- 26. The method of claim 25, wherein the soluble CTLA4 mutant molecule comprises an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 27. The method of claim 25, wherein the soluble CTLA4 mutant comprises an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 28. The method of claim 25, wherein said T cell interactions are inhibited.
- 29. A method for inhibiting graft versus host disease in a subject which comprises administering to the subject the soluble CTLA4 mutant molecule of claim 1 and a ligand reactive with IL-4.
- 30. The method of claim 29, wherein the soluble CTLA4 mutant molecule comprises an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 31. The method of claim 29, wherein the soluble CTLA4 mutant comprises an extracellular domain of CTLA4, wherein in the extracellular domain, alanine at position +29 is substituted with tyrosine and leucine at position +104 is substituted with glutamic acid as shown in FIG. 7.
- 32.
Parent Case Info
[0001] This application is a continuation-in-part of U.S. Ser. No. 09/014,761, filed Jan. 28, 1998, which claims priority of U.S. Serial No. 60/036,549, filed Jan. 31, 1997, now abandoned, the contents of all of which are incorporated by reference into the present application.
Provisional Applications (1)
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Number |
Date |
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60036594 |
Jan 1997 |
US |
Continuations (1)
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Number |
Date |
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Parent |
09603825 |
Jun 2000 |
US |
Child |
10336384 |
Jan 2003 |
US |
Continuation in Parts (1)
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Number |
Date |
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Parent |
09014761 |
Jan 1998 |
US |
Child |
09603825 |
Jun 2000 |
US |