Establishing Swine To Monkey Disparate Hematopoietic Chimera And Changing Of Coagulation In The Model

Clinical homeotransplantation has developed rapidly, but shortage of donor organs has become restriction of organ transplantation development. Thus, with rapid development of transplantation rejection molecular theory and cellular theory, heterogeneous transplantation is a valid method to solve the problem. The cultivation of transgenic animal with human gene and to establish donor-recipient hemopoietic chimera for immunotolerance has bright applied prospects. We have established rat to rabbit heterogeneous hemopoietic chimera model successfully. In the model, rat's mononuclear cells were chimerized in the recipient over a long-term while recipient's antibody and complement exists and their functions are normal. The results encouraging us badly, but it is established based on small animals, it's far from used in clinical actually. So we should find bigger mammal that close to human used for research xenotransplantation. Pig is known that it is most suitable donor for xenotransplantation to-6-human. So we design the subject to select pig and monkey as donor and recipient for xenotransplantation research.Aim:By transplanting pig's thymus and marrow into monkeys, the pig's immunosystem is set up in the monkeys so as to induce recipients' immunotolerance to the donors. While oberserve changes of coagulopathy in recipients.Methods:1. Pretreatment of recipent monkeys: recipients, adult monkeys, were splenectomized and then treated with cyclophosphamide2-4 times with some immuno inhibitor(MMF and CsA).2. Transplantation of pig's thymus and marrow: Thymus tissue pieces of pigs were transplanted into monkeys' neck and back under skin. Suspension of pig's marrow cells was injected into recipients monkeys through vein. After transplantation, inject ALG, MMF and CsA.3. Determination: Through PCR, we examined the pig's special sequence in DMA of recipients monkeys' peripheral blood mononuclear cells and through Wenstern blot to find recipients monkeys' peripheral blood mononuclear cells express pig's MHC I antigen. Observe recipent's pathology change through pathological examination.4. Through blood examnation to observe recipent's coagulopathy changing.Results:1. 2 monkeys dead after transplantation and 4 monkeys lived long time. And we found that there are some GVHD symptom among recipients just like lose weight, low energy and no interesting to food. Blood examination support the symptom.2. PCR examnation and Western blot conformed that the modle of heterogeneous hemoporetic chimera was established. And donor's cells and recipient's cells chimera together could last for 30 days.3. After tranplantation, all recipients occured coagulopathy. That is levels of Platelet count and Fibrinogen decreased, but prothrombin time, level of D-Dimers and TAT increased. And 2 dead monkey occued DIG.Conclusions:1. PCR examnation and Western blot conformed that the modle of heterogeneous hemoporetic chimera was established. So we could say it is successful method for establish swine to monkey disparate hematopoietic chimera ?2. Both blood and pathological examnation only conform GVHD is not severe. So the method for establish swine to monkey disparate hematopoietic chimera is safe.3. After tranplantation, all recipients occured coagulopathy. And 2 dead monkey occued DIG. That we could say DIG may be third barrier to xenotransplantation besides humoral and cell rejection.4. The study is short relatively for chimera. We think because there is short of cytokine for supporting donor's bone marrow cells and other cells growing in recipients body.