Porcine CTLA-4 for xenograft-specific immunosuppression
The invention provides methods for inhibiting T-cell mediated rejection of a xenotransplanted organ by blocking the delivery of co-stimulatory signal 2 (the B7/CD28 interaction) prevent the activation of xenoreactive T-cells in the recipient. In a first aspect, co-stimulation is prevented by administration to the organ recipient of a soluble form of CTLA-4 from the xenogeneic donor organism. In a second aspect, co-stimulation is antagonized by expressing a ligand for CTLA-4 on the xenogeneic donor cells. In a third aspect, co-stimulation is prevented by expressing recipient organism MHC class II on the surface of the cells of the xenogeneic donor organ.
1. A non-human cell transformed with a vector which expresses a nucleic acid which encodes a porcine CTLA4 protein or fragment thereof.
2. The cell according to claim 1 wherein the CTLA4 protein comprises the extracellular domain of porcine CTLA-4.
3. The cell according to claim 1 , wherein the CTLA-4 protein comprises the extracellular domain of porcine CTLA4 fused to a human Cγ1 sequence.
4. The cell of claim 1 wherein the nucleic acid comprises SEQ ID NO: 2.
5. The cell according to claim 1 wherein the CTLA4 protein is fused to the constant domain of an immunoglobulin.
6. The cell of claim 3 , wherein the immunoglobulin gamma comprises a constant region of the human Cγ1 subtype.
7. The cell of claim 5 , wherein a linker connects the porcine CTLA-4 to the immunoglobulin and the linker comprises the amino acid sequence GGSGGAA (SEQ ID NO: 28).
8. The cell of claim 1 wherein the CTLA-4 protein is fused to an immunoglobulin.
9. The cell of claim 8 , wherein the immunoglobulin is a human immunoglobulin.
10. The cell of claim 8 , wherein the human immunoglobulin is immunoglobulin gamma (IgG).
11. The cell of claim 10 , wherein the immunoglobulin gamma comprises a constant region of the human Cγ1 subtype.
12. The cell of claim 8 , wherein a linker connects the porcine CTLA-4 to the immunoglobulin.
13. The cell of claim 12 , wherein the linker comprises the amino acid sequence GGSGGAA (SEQ ID NO: 28).
14. The cell of claim 2 wherein the extracellular domain of the porcine CTLA-4 protein comprises amino acid sequence numbers 38-161 of SEQ ID NO: 1.