Transgenic ungulates expressing CTLA4-IG and uses thereof
The present invention provides ungulates, including pigs, expressing CTLA4-Ig, as well as tissue, organs, cells and cell lines derived from such animals. Such animals, tissues, organs and cells can be used in research and medical therapy, including xenotransplanation. In addition, methods are provided to prepare organs, tissues and cells expressing the CTLA4-Ig for use in xenotransplantation, and nucleic acid constructs and vectors useful therein.
1. A method of reducing cell mediated rejection of xenotransplanted cells in a recipient, comprising xenotransplanting genetically modified porcine cells into the recipient wherein:
(i) the cell mediated rejection of the genetically modified porcine cells is reduced without suppression of the recipient's normal immune function;
(ii) the genetically modified cells lack expression of the alpha-1,3-galactosyltransferase (alpha-1,3-GT) gene and express a transgene encoding a porcine CTLA4 peptide fused to a human immunoglobulin peptide (pCTLA4-Ig); and
(iii) the genetically modified cells are directly obtained from a genetically modified porcine animal that lacks expression of the alpha-1,3-GT gene and expresses the pCTLA4-Ig peptide by insertion of the pCTLA4-Ig transgene into the genome of the animal.
2. The method of claim 1 , wherein the CTLA4 peptide is truncated.
3. The method of claim 1 , wherein the CTLA4 peptide is mutated.
4. The method of claim 1 , wherein the CTLA4 peptide is mutated by substitution of (a) an alanine at position +29 with a tryptophan, and (b) a leucine at position +104 with a glutamic acid.
5. The method of claim 1 , wherein the CTLA4 peptide is modified by the addition of an intracellular retention signal.
6. The method of claim 1 , wherein the Ig is IgG.
7. The method of claim 1 , wherein the IgG is IgG1 or IgG4.
8. The method of claim 1 , wherein the cells are obtained from a tissue or organ of the porcine of claim 1 .
9. The method of claim 1 , further comprising administering soluble CTLA4 in addition to the genetically modified cells to the recipient.
10. The method of claim 9 , wherein the soluble CTLA4 is porcine.
11. The method of claim 9 , wherein the soluble CTLA4 is human.