Surrogate tolerogenesis for the development of tolerance to xenografts

• US 6,060,049 A
• Filed: 06/06/1995
• Issued: 05/09/2000
• Est. Priority Date: 05/24/1993
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Ximerex Incorporated
• Inventors: Beschorner, William E.
• Primary Examiner(s): Crouch, Deborah
• Assistant Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US08/295,899
• Time in Patent Office: 1,799 Days
• Field of Search: 424/529, 424/577, 424/93.7, 424/93.21, 424/93.71, 424/2, 424/9, 424/93.3, 435/1, 435/240.2, 435/375, 435/377, 800/DIG. 5, 800/2, 800/8
• US Class Current: 424/93.21
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 35/26 Lymph; Lymph nodes; Thymus; Spleen; Splenocytes; Thymocytes
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 39/001 Preparations to induce tolerance to non-self, e.g. prior to transplantation
• CPC Class Codes: A61K 39/461 characterised by the cell type used
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/46434 Antigens related to induction of tolerance to non-self
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection

Abstract

This invention provides a method for developing immune tolerance in xenogeneic organ graft recipients, in which lympho-hematopoietic cells from an intended organ graft recipient are differentiated within a xenogeneic surrogate, such as a fetal animal. After birth of the surrogate, the matured lympho-hematopoietic cells containing antigen specific regulatory cells, including suppressor cells, veto cells, select B cells, anti-idiotype antibodies, and other related factors responsible for antigen specific tolerance in a surrogate animal are reintroduced into the intended organ graft recipient, in conjunction with an organ transplant or a tissue transplant from the xenograft surrogate. The invention also provides an organ graft repopulated with cells from the intended organ graft recipient produced in a surrogate animal.

Claims

1. A method for xenograft transplant of an organ to a recipient mammal from a donor mammal, wherein said recipient has increased immuno tolerance to donor tissue, comprising the steps of:

a) collecting a first cell population from the recipient, said first cell population containing lymphocytic progenitor cells, wherein at least a portion of cells that are specifically cytotoxic to tissue from a surrogate animal are removed from said first cell population;

b) administering said first cell population to said surrogate, said surrogate being in a state of immune deficiency;

c) developing in said surrogate a state of immune competence;

d) collecting from said immune competent surrogate a second population of cells, said second cell population containing immunocompetent cells, said immunocompetent cells specifically suppressing immune response of said recipient to tissue of said surrogate;

e) infusing said second population of cells into said recipient;

f) excising an organ from a donor mammal wherein the donor mammal is selected from the group consisting of the same surrogate mammal, the same inbred strain as the surrogate mammal and a littermate of the surrogate mammal; and

g) transplanting said organ into said xenograft recipient.

2. The method of claim 1, wherein said first cell population from the recipient comprises bone marrow cells.

3. The method of claim 1, wherein said surrogate is a fetal mammal or a neonatal mammal.

4. The method of claim 1 further comprising treating said recipient with an immunosuppressive agent before transplanting said organ into said xenograft recipient.

5. The method of claim 1, wherein said second population of cells comprises spleen cells.

6. The method of claim 1, wherein the organ from the donor animal comprises vascular tissues.

7. The method of claim 1 wherein the said donor mammal is the same inbred strain as the surrogate mammal.

8. The method of claim 1 wherein the said donor mammal is a littermate of the surrogate mammal.

9. A method of transplanting an organ from a donor mammal to a recipient mammal which is not syngeneic with said donor mammal, comprising

a) administering to said recipient mammal a cell population containing immunocompetent cells specifically suppressing immune response of said recipient mammal to said donor mammal, wherein said cell population is obtained from a surrogate mammal, said surrogate mammal being a chimeric animal containing lymphocytes derived from said recipient mammal;

and

b) transplanting an organ from said donor mammal to said recipient mammal, wherein said donor mammal is selected from the group consisting of the same surrogate mammal, the same inbred strain as the surrogate mammal, and a littermate of the surrogate mammal,

whereby immune response of said recipient mammal to said organ is reduced as compared to the immune response of said recipient mammal in the absence of the administration of said surrogate mammal cell population.

10. The method of claim 9, wherein said surrogate mammal and said recipient mammal are from different species.

11. The method of claim 10, wherein said surrogate mammal and said donor mammal are different individual mammals.

12. The method of claim 10, wherein said donor mammal and said recipient mammal are from the same species.

13. The method of claim 9, wherein said donor mammal is mature when said surrogate mammal is born.

14. The method of claim 9, wherein said donor mammal and said surrogate mammal are different individuals and said organ is a population of cells, further wherein said cell population is expanded by culturing in said surrogate mammal before transplanting into said recipient mammal.

15. The method of claim 14, wherein said organ is selected from the group consisting of pancreatic islet cells and hepatic cells.

16. The method of claim 14, wherein said organ is bone marrow.

17. A method of transplanting an organ from a third party donor mammal to a recipient mammal which is of the same species as said third party donor mammal, comprising

a) administering to said recipient mammal a cell population containing immunocompetent cells specifically suppressing immune response of said recipient mammal to said third party donor mammal, wherein said cell population is obtained from a surrogate mammal, said surrogate mammal being a chimeric mammal containing lymphocytes derived from a set of mammals of the same species as the recipient, said set of mammals having a plurality of tissue types;

and

b) transplanting an organ from said third party donor mammal to said recipient mammal,

whereby immune response of said recipient mammal to said organ is reduced as compared to the immune response of said recipient mammal in the absence of the administration of said surrogate mammal cell population.

40. The method of claim 39, further comprising, before the step of collecting said cell population of claim 17, the steps of:

i) collecting respective blood or spleen samples from the plurality of immune competent developed surrogates;

ii) testing the respective blood samples to determine a degree of engraftment with cells of said recipient mammal and to determine a degree of maturation of lymphocytes in the respective blood samples;

iii) testing respective cells of the plurality of developed surrogates for specific suppression of immune reactivity between recipient mammal cells and donor mammal cells;

iv) selecting, in response to the steps of testing the respective blood samples, a selected set of the plurality of developed surrogates, said selected set having the greatest degree of engraftment and the greatest immune suppression;

v) testing for graft-vs-host disease (GvHD) against respective developed surrogates of the selected set by collecting fresh lymphocytes from said xenograft recipient and infusing said fresh lymphocytes into the selected set of developed surrogates; and

vi) identifying a best developed surrogate from the selected set of developed surrogates, based on least degree of GvBD in response to infusion of lymphocytes from the xenograft recipient.

18. A kit for suppression of immune rejection by a recipient mammal of an organ transplanted from a third party donor mammal which is of the same species as the surrogate mammal comprising an immune suppressive composition comprising a cell population obtained from a surrogate mammal, said surrogate mammal being a chimeric mammal containing lymphocytes derived from said recipient mammal and expressing antigens substantially identical to said third party donor mammal, and said cell population containing immunocompetent cells specifically suppressing immune response of said recipient mammal to said third party donor mammal, said cell population being suspended in a medium suitable for injection into said recipient mammal.

19. The kit of claim 18, wherein said immune suppressive composition is substantially free of immune reactive cells of said surrogate.

20. The kit of claim 18, wherein the composition suitable for injection into said recipient mammal is free of infectious agents.

21. A kit for organ transplant into a recipient mammal comprising an excised bodily organ from a surrogate mammal wherein said surrogate is different from said recipient and perfusion solution in an amount sufficient to preserve said excised organ in condition suitable for transplant into said recipient mammal, wherein a plurality of resident cells of said organ are cells from the same species as said recipient, said resident cells being selected from endothelial cells, monocytes, dendritic cells, and lymphoid cells, said bodily organ being placed in the perfusion solution and preserved in condition suitable for transplant into said recipient mammal.

22. The kit of claim 21, wherein said plurality of resident cells are syngeneic with said recipient mammal.

23. A method of preparing an excised organ of claim 21 for transplant into a recipient mammal which is not syngeneic with said excised organ, comprising:

a) collecting a cell population from said recipient mammal, said first cell population containing lymphocytic progenitor cells;

b) administering said cell population to a surrogate mammal, said surrogate being in a state of immune deficiency;

c) developing in said surrogate a state of immune competence;

d) excising from said immune competent surrogate an organ, said organ being populated with at least a plurality of cells derived from said recipient mammal; and

e) placing said organ in perfusion solution so that said excised organ is preserved in condition suitable for transplant into said recipient mammal.

24. The method of claim 23, further wherein at least a portion of cells that are specifically cytotoxic to tissue from said surrogate mammal are removed from said cell population before said cell population is administered to said surrogate.

25. A method of preparing an excised organ for transplant into a recipient mammal which is not syngeneic with said excised organ, comprising:

a) collecting a cell population from an animal which is of the same species as said recipient mammal, said cell population containing lymphocytic progenitor cells;

b) administering said cell population to a surrogate mammal, said surrogate being in a state of immune deficiency;

c) developing in said surrogate a state of immune competence;

d) excising from said immune competent surrogate an organ, said organ being populated with a plurality of cells derived from said recipient mammal; and

e) placing said organ in perfusion solution so that said excised organ is preserved in condition suitable for transplant into said recipient mammal.

26. The method of claim 25, further wherein at least a portion of cells that are specifically cytotoxic to tissue from said surrogate mammal are removed from said cell population before said cell population is administered to said surrogate.

27. The method of claim 25, wherein lymphocytes derived from a set of mammals of the same species as the recipient are administered to said surrogate mammal while it is in a state of immune deficiency, said set of mammals having a plurality of tissue types.

28. The method according to claim 27, wherein said recipient mammal is a human having severe burns, said surrogate mammal is a non-human mammal, and said organ is a graft of skin from said non-human mammal for application to the burned area of said recipient, wherein a plurality of resident cells of said skin are cells derived from the same species as said patient.

29. The method of claim 25, wherein said donor mammal and said surrogate mammal are different individuals and said organ is a population of cells, further wherein said cell population is expanded by culturing in said surrogate mammal before transplanting into said recipient mammal.

30. The method of claim 29, wherein said organ is selected from the group consisting of pancreatic islet cells and hepatic cells.

31. The method of claim 29, wherein said organ is bone marrow.

32. A method of therapy for a patient having fulminant hepatitis, comprising periodically transplanting hepatic cells prepared according to claim 30 into said patient so long as hepatic necrosis continues.

33. A method of suppressing specific immune response of a recipient human to a transplantable organ of a non-human mammal comprising administering to the recipient human, prior to transplantation of the organ of a non-human mammal, a cell population containing immunocompetent cells specifically suppressing immune response of the recipient human to the organ of the non-human mammal, the cell population being obtained from a non-human surrogate mammal, the non-human surrogate mammal being a chimeric animal containing said immunocompetent cells specifically suppressing human immune response to cells from the non-human mammal, the said immunocompetent cells being derived from the recipient human, whereby immune response of the recipient human to the organ is reduced as compared to the immune response of said recipient mammal in the absence of the administration of said surrogate mammal cell population.

34. The method of claim 33, further wherein at least a portion of cells that are specifically cytotoxic to tissue from said surrogate mammal are removed before cells are administered to said surrogate.

35. The method of claim 33, further comprising, before the step of collecting said cell population, a step of collecting fresh lymphocytes from said recipient and infusing said fresh lymphocytes into said developed surrogate.

36. The method of claim 33, wherein said surrogate is a fetal mammal.

37. The method of claim 33, wherein the surrogate is selected from the group consisting of a newborn mammal, a juvenile mammal and an adult mammal, said mammal having received lethal irradiation or chemotherapy followed by bone marrow transplant of a cell population comprising hematopoietic and lymphoid cells from an organism selected from said recipient mammal, said donor mammal, and said surrogate, or a mixture of hematopoietic and lymphoid cells from two or more of these organisms.

38. The method of claim 33, wherein said surrogate is the same mammal as said donor mammal.

39. The method of claim 33, wherein said lymphocytes are additionally administered to a plurality of immune deficient surrogates and said plurality of surrogates is allowed to develop into immune competent organisms.

41. The method of claim 33, wherein lymphocytes derived from both genetic parents of said donor mammal are administered to said surrogate mammal while it is in a state of immune deficiency.

42. The method of claim 41, wherein said organ is a cell suspension, and further wherein the cell suspension contains cells from more than one off-spring of said genetic parents.

43. The method of claim 41, wherein said donor mammal is mature when said surrogate mammal is born.

44. The method of claim 33, wherein said donor mammal and said surrogate mammal are different individuals and said organ is a cell suspension, further wherein said cell suspension is expanded by culturing in said surrogate mammal before transplanting the expanded cell suspension into said recipient mammal.

45. The method of claim 44, wherein said organ is selected from the group consisting of pancreatic islet cells and hepatic cells.

46. The method of claim 44, wherein said organ is bone marrow.

47. The method of claim 33, wherein the immunocompetent cells are human lymphocytes derived from hematopoietic cells of the recipient human introduced into the surrogate animal when the surrogate mammal was in a state of immune deficiency, the immunocompetent cells being collected from the surrogate mammal after the surrogate mammal achieved a state of immune competency.

48. The method of claim 33, wherein the non-human surrogate mammal is a primate, an artiodactyl or a carnivore.

49. The method of claim 33, wherein the non-human surrogate mammal is a primate or a pig.

50. The method of claim 33, wherein the non-human surrogate mammal is a rodent or a lagomorph, and the transplantable organ is a suspension of cells.

51. A method of preparing an excised non-human organ suitable for transplant into a recipient human comprising:

a) collecting a human cell population containing lymphocytic progenitor cells;

b) administering the cell population to a non-human donor mammal, the mammal being in a state of immune deficiency;

c) developing in the non-human mammal a state of immune competence;

d) excising an organ from the immune competent non-human mammal, the organ being populated with a plurality of cells derived from the human cell population; and

e) placing the organ in perfusion solution so that the excised organ is preserved in condition suitable for transplant into a recipient human.

52. The method of claim 51, wherein the non-human mammal is a primate, an artiodactyl or a carnivore.

53. The method of claim 51, wherein the non-human surrogate mammal is a primate or a pig.

54. The method of claim 53, wherein the non-human mammal in a state of immune deficiency is a fetal mammal.

55. The method of claim 51, wherein the human cell population is obtained from the recipient human.