Combined organ and hematopoietic cells for transplantation tolerance of grafts
Methods and compositions are provided for combined transplantation of a solid organ and hematopoietic cells to a recipient, where tolerance to the graft is established through development of a persistent mixed chimerism. An individual with persistent mixed chimerism, usually for a period of at least six months, is able to withdraw from the use of immunosuppressive drugs after a period of time sufficient to establish tolerance.
1. A method for manufacturing a cryopreserved cellular product for establishing mixed chimerism in a solid organ transplant recipient, the method comprising:
obtaining greater than 5×10 5 CD34+ cells/kg recipient weight;
obtaining greater than 1×10 5 CD3+ cells/kg recipient weight;
cryopreserving the greater than 5×10 5 CD34+ cells/kg recipient weight in a first cryopreservation solution comprising DMSO at a substantially neutral pH; and
cryopreserving the greater than 1×10 5 CD3+ in a second cryopreservation solution comprising DMSO at a substantially neutral pH.
2. The method of claim 1 , wherein the CD34+ cells and the CD3+ cells are cryopreserved in separate containers.
3. The method of claim 1 , wherein the CD34+ cells and the CD3+ cells are cryopreserved as a mixture in a common container.
4. The method of claim 1 , wherein at least one of the first cryopreservation solution and the second preservation solution comprises a polymeric carbohydrate.
5. The method of claim 1 , wherein the first cryopreservation solution and the second preservation solution are different.
6. The method of claim 1 , wherein the first cryopreservation solution and the second preservation solution are the same.
7. The method of claim 1 , wherein the CD34+ cells and the CD3+ cells are HLA matched to an HLA type of the solid organ transplant recipient.
8. The method of claim 1 , wherein the CD34+ cells and the CD3+ cells are HLA mismatched to an HLA type of the solid organ transplant recipient.
9. The method of claim 1 , wherein the CD34+ cells and the CD3+ cells are from a donor related to the solid organ transplant recipient.
10. The method of claim 1 , wherein CD34+ cells and the CD3+ cells are from a single apheresis product.
11. A method for manufacturing a cryopreserved cellular product for establishing mixed chimerism in a solid organ transplant recipient that is HLA matched to the solid organ transplant donor, the method comprising:
obtaining greater than 5×10 5 CD34+ cells/kg recipient weight, wherein the solid organ transplant donor's CD34+ cells are of an HLA type that is matched to the solid organ transplant recipient's HLA type;
obtaining a defined amount of about 1×106 CD3+ cells/kg recipient weight, wherein the solid organ transplant donor's CD3+ cells are of an HLA type that is matched to the solid organ transplant recipient's HLA type; and
cryopreserving the greater than 5×10 5 CD34+ cells/kg recipient weight and the defined amount of about 1×106 CD3+ cells/kg recipient weight in a cryopreservation solution comprising DMSO.
12. The method of claim 11 , wherein the CD34+ cells and the CD3+ cells are cryopreserved as a mixture in a common container comprising the cryopreservation solution.
13. The method of claim 11 , wherein the cryopreservation solution comprises a polymeric carbohydrate.
14. The method of claim 11 , wherein the cryopreservation solution comprises less than 10% DMSO.
15. The method of claim 11 , wherein the product is at a substantially neutral pH.
16. The method of claim 11 , wherein the CD34+ cells and the CD3+ cells are HLA matched to the solid organ transplant recipient at all six alleles of HLA-A, HLA-B, and HLA-DR.
17. The method of claim 11 , wherein the CD34+ cells and the CD3+ cells are from a donor related to the solid organ transplant recipient.
18. The method of claim 11 , wherein CD34+ cells and the CD3+ cells are from a single apheresis product.
19. The method of claim 11 , wherein CD34+ cells and the CD3+ cells are from multiple apheresis products.
20. The method of claim 11 , wherein the solid organ is selected from the group consisting of heart, intestine, liver, lung, pancreas, and kidney.
21. A method for manufacturing a cryopreserved cellular product for establishing mixed chimerism in a solid organ transplant recipient that is HLA mismatched from the solid organ transplant donor, the method comprising:
obtaining greater than 5×10 5 CD34+ solid organ transplant donor derived cells/kg recipient weight, wherein the solid organ transplant donor's CD34+ cells are of an HLA type that is mismatched to the solid organ transplant recipient's HLA type;
obtaining greater than 4×10 7 CD3+ solid organ transplant donor derived cells/kg recipient weight, wherein the solid organ transplant donor's CD3+ cells are of an HLA type that is mismatched to the solid organ transplant recipient's HLA type;
cryopreserving the greater than 5×10 5 CD34+ solid organ transplant donor derived cells/kg recipient weight in a first cryopreservation solution comprising DMSO in a first vessel; and
cryopreserving the greater than 4×10 7 CD3+ solid organ transplant donor derived cells/kg recipient weight in a second cryopreservation solution comprising DMSO in a second vessel.
22. The method of claim 21 , wherein at least one of the first cryopreservation solution and the second preservation solution comprises a polymeric carbohydrate.
23. The method of claim 21 , wherein the first cryopreservation solution and the second preservation solution are different.
24. The method of claim 21 , wherein the first cryopreservation solution and the second preservation solution are the same.
25. The method of claim 21 , wherein each of the first cryopreservation solution and the second cryopreservation solution comprises less than 10% DMSO.
26. The method of claim 21 , wherein the CD34+ cells and the CD3+ cells are HLA mismatched to the solid organ transplant recipient at at least one of six alleles of HLA-A, HLA-B, and HLA-DR.
27. The method of claim 1 , wherein the product is at a substantially neutral pH.
28. The method of claim 21 , wherein CD34+ cells and the CD3+ cells are from a single apheresis product.
29. The method of claim 21 , wherein CD34+ cells and the CD3+ cells are from multiple apheresis products.
30. The method of claim 21 , wherein the solid organ is selected from the group consisting of heart, intestine, liver, lung, pancreas, and kidney.