Inducement of organogenetic tolerance for pancreatic xenotransplant
Provided herein is an approach to establish organogenetic tolerance via prior transplantation of pig embryonic pancreas, thereby enabling subsequent implantation of porcine islets in a subject without the need for immune-suppression. In one aspect of the invention, porcine pancreatic primordia are implanted into a mammalian subject, and after a period of time sufficient to induce tolerance, porcine islet cells are implanted in the subject.
1. A method for treating diabetes mellitus in a subject in need thereof comprising:
implanting a porcine pancreatic primordium, or a portion thereof, in an amount effective to induce tolerance to porcine islet cells comprising mature antigen-presenting cells, into a non-porcine mammalian subject;
waiting a period of time sufficient to induce tolerance to porcine islet cells comprising mature antigen-presenting cells; and
implanting porcine islet cells comprising mature antigen-presenting cells into the non-porcine mammalian subject.
2. The method of claim 1 wherein a requirement for immunosuppression is reduced or eliminated.
3. The method of claim 1 , wherein the porcine pancreatic primordium is isolated about 7 days after formation of an embryonic pig pancreas.
4. The method of claim 1 , wherein the developmental age of the porcine pancreatic primordium is at least about E27 but not more than about E35.
5. The method of claim 4 , wherein the developmental age of the porcine pancreatic primordium is about E28.
6. The method of claim 1 , wherein the pancreatic primordium comprises at least one embryonic dorsal, ventral, or fused pancreatic primordium that is substantially non-vascularized at the time of the harvest.
7. The method of claim 1 , wherein
the pancreatic primordium comprises at least one embryonic dorsal, ventral, or fused pancreatic primordium; and
the pancreatic primordium is substantially free of antigen-presenting cells at the time of the harvest.
8. The method of claim 1 , wherein at least about one pancreatic primordium, or an equivalent amount of portions of a pancreatic primordium sufficient to induce tolerance to porcine islet cells, are implanted into the subject.
9. The method of claim 8 , wherein at least about 1 to about 20 pancreatic primordia, or an equivalent amount of portions of pancreatic primordia sufficient to induce tolerance to porcine islet cells, are implanted into the subject.
10. The method of claim 1 , wherein the pancreatic primordia are implanted at one or more positions selected from the group consisting of: peritoneal cavity; mesentery; near the subject's omentum adjacent to a branch of the subject's superior mesenteric artery; into a pouch of the omentum; in the subject's kidney capsule; under the subject's kidney capsule; within skeletal muscle; and in the subcutaneous space.
11. The method of claim 1 , wherein the period of time sufficient to induce tolerance to porcine islet cells is at least about 10 days after implantation of the porcine pancreatic primordia.
12. The method of claim 1 , wherein the implanted porcine pancreatic primordia remains in place in the subject during the period of time sufficient to induce tolerance to porcine islet cells.
13. The method of claim 1 , wherein the porcine islet cells are implanted into the subject in an amount of at least about 10,000 islet equivalents per kg of the subject.
14. The method of claim 1 , wherein the porcine islet cells are implanted at one or more sites by intrahepatic islet infusion or intraportal injection.
15. The method of claim 1 , wherein the porcine islet cells comprise at least one of: α cells; β cells; δ cells; PP cells; and stem cells.
16. The method of claim 1 , further comprising monitoring the subject for one or more of: implanted islet cell function; islet cell graft loss; insulin secretion, hematological and biochemical parameters; and plasma levels of porcine insulin.
17. The method of claim 1 , wherein the mammalian subject is selected from the group consisting of: horse, cow, dog, cat, sheep, pig, mice, rat, monkey, guinea pig, chicken, and human.
18. The method of claim 17 , wherein the mammalian subject is a human.
19. The method of claim 1 , wherein the diabetes mellitus comprises at least one of: Type 1 diabetes, Type 2 diabetes, pre-diabetes, gestational diabetes, congenital diabetes due to genetic defects of insulin secretion, cystic fibrosis-related diabetes, steroid diabetes induced by high doses of glucocorticoids, and monogenic diabetes.
20. The method of claim 19 , wherein the diabetes mellitus is Type 1 diabetes or Type 2 diabetes.
21. The method of claim 1 , wherein the porcine islet cells are implanted at one or more sites selected from the group consisting of: in the peritoneal cavity; in subcutaneous tissue; and in the omentum of the subject.
22. The method of claim 1 , wherein the porcine islet cells are implanted at one or more sites selected from the group consisting of: in the kidney capsule; underneath the kidney capsule; in epididymal fat; and in pancreas of the subject.