Methods of treatment using CTLA4 mutant molecules
The present invention provides soluble CTLA4 mutant molecules which bind with greater avidity to the CD80 and/or CD86 antigen than wild type CTLA4 or non-mutated CTLA4Ig. The soluble CTLA4 molecules have a first amino acid sequence comprising the extracellular domain of CTLA4, where certain amino acid residues within the S25-R33 region and M97-G107 region are mutated. The mutant molecules of the invention may also include a second amino acid sequence which increases the solubility of the mutant molecule.
1. A method for treating transplant rejection in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule binds CD80 and/or CD86 and comprises the extracellular domain of CTLA4 as shown in SEQ ID NO:8 beginning with alanine at position 26 or methionine at position 27 and ending with aspartic acid at position 150, or a portion thereof that binds CD80 and/or CD86, wherein in the extracellular domain or portion thereof an alanine at position 55 is substituted with a tyrosine, and a leucine at position 130 is substituted with a glutamic acid.
2. A method for inhibiting graft versus host disease in a subject which comprises administering to the subject a CTLA4 mutant molecule and a ligand reactive with IL-4, wherein the CTLA4 mutant molecule comprises:
(a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4.
3. The method of claim 2 , wherein the CTLA4 mutant molecule comprises:
(a) an amino acid sequence beginning with methionine at position 27 and ending with lysine at position 383 of SEQ ID NO:4, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with lysine at position 383 of SEQ ID NO:4.
4. A method for treating transplant rejection in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
(a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4.
5. A method for treating transplant rejection in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
(a) an amino acid sequence beginning with methionine at position 27 and ending with aspartic acid at position 150 of SEQ ID NO:4 or a portion thereof that binds CD80 and/or CD86, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with aspartic acid at position 150 of SEQ ID NO:4 or a portion thereof that binds CD80 and/or CD86.
6. The method of claim 4 , wherein the CTLA4 mutant molecule further comprises an amino acid sequence which alters the solubility or affinity of the CTLA4 mutant molecule.
7. The method of claim 6 , wherein the amino acid sequence which alters the solubility or affinity comprises an immunoglobulin.
8. The method of claim 7 , wherein the immunoglobulin is an immunoglobulin constant region or portion thereof.
9. The method of claim 8 , wherein the immunoglobulin constant region or portion thereof is mutated to reduce effector function.
10. The method of claim 8 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule.
11. The method of claim 9 , wherein the immunoglobulin constant region or portion thereof comprises a hinge, CH2 and CH3 regions of a human or monkey immunoglobulin molecule.
12. A method for treating transplant rejection in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule comprises:
(a) an amino acid sequence beginning with methionine at position 27 and ending with lysine at position 383 of SEQ ID NO:4, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with lysine at position 383 of SEQ ID NO:4.
13. A method for treating transplant rejection in a subject comprising administering to the subject a CTLA4 mutant molecule, wherein the CTLA4 mutant molecule consists of:
(a) an amino acid sequence beginning with methionine at position 27 and ending with lysine at position 383 of SEQ ID NO:4, or
(b) an amino acid sequence beginning with alanine at position 26 and ending with lysine at position 383 of SEQ ID NO:4.
14. A method for treating transplant rejection in a subject comprising administering to the subject the CTLA4 mutant molecule encoded by the nucleic acid molecule designated ATCC No. PTA-2104.
15. The method of claims 4 , 8 , 9 , 10 , 11 , 12 , or 14 wherein the transplant rejection is selected from the group consisting of solid organ and/or tissue transplant rejection, allo- or xenograft acute or chronic rejection, tissue or cell allo- or xenograft rejection, skin transplant rejection, islet transplant rejection, muscle transplant rejection, hepatocyte transplant rejection, and neuron transplant rejection.
16. The method of claim 15 , wherein the CTLA4 mutant molecule is administered concomitantly or in sequence with one or more agents selected from the group consisting of corticosteroids, azathioprine, tacrolimus, basiliximab, cyclosporin A, rapamycin, 40-O-(2-hydroxy)ethyl-rapamycin, fingolimod; cyclophosphamide; methotrexate; leflunomide; mizoribine; mycophenolic acid; mycophenolate mofetil; 15-deoxyspergualine, and CTLA4/CD28-lg.