| Graft versus host disease (GVHD) is caused by immune competent cells -mostly T cells, which recognize the host's antigens and hurt the host? tissues. Deleting or killing the T cells in the allograft will certainly reduce the morbidity of GVHD. from 80 percent to 30 percent. but at the same time, will also contribute to the relapse of leukemia(by 30 percent). the failure of transplantation(by 20 percent) and infection(by 30 percent). More and more research estimate that immunocompetent cells in the allograft will help a lot to the reconstitution of immunosystem and blood making. A new promising orientation is to decrease the transplantation related complications and improve the effect of it through mediating T cell's function. Subject Using the polarized Th2 Tc2 cell transplantation to establish a new model for bone marrow transplantation, reducing GVHD but retaining GVL effect. Discussing the detachment of GVHD and GVL from the molecular mechanism of the recognization of KIRIMHC I to lay a foundation for GVHD research Methods 1. Production and detection of Th2 Tc2 polarized cells. In vitro using mIL-4. ionomycine and conA to education the C57BL/6 mice T cells to turn toward Th2 Tc2 cells. Using ELISA and FCM to make sure that the cells producing IL-4. 2. Detection of expression and function of Ly49c Ly49A on the surface of Th2 Tc2. 3. Establishment of mice model of acute GVHD after haploidentical allogenetic bone marrow transplantation: the donor is C57BL/62b. The recipient is BalB/cxB6 Fl H2d/b(CB6F1) After the radiation of (TBI, 60Co) the recipient accept the transplantation mixed T cell and bone marrow cells to establish the acute GVHD model. 4. Establishment of erythroleukemia model of halploidentical mice. Transfer the erythroleukemia cell EL961 1H-2d the CB6F1 I]-2d/b mice to H 2d/b establish the Fl - erythroleukemia model providing the test animal model for detecting GVL effect after halploidentical Th2 Tc2 cell transplantation. 5. Detection of GVHD GVL effect after the transplantation of bone marrow cells mixed with Th2 Tc2 cells: themixture of Th2 Tc2 cells with the bone marrow cells from C57BL/6 H 2bmice at the ratio of 10:1 (polaried T cells 5 x 1 0, bone marrow cells 5 x 106) and were transfused into the predisposed haploidentical nomoral or erythroleukemia recipient mice CB6F I. Using survival time and pathological analysis to estimate the transplantation effect. At the same time, detecting the expression of Ly49A Ly49C to discuss the relationship between KIR expression and GVHD,GVL. Results 1. The spleen T lymphoxcytes can be induced to class II cytokines producing cells in vitro after 4 days culturing with mIL-4 ConA and Ionomycin. The mIL-4 secreating level rises from 20 I.1 g/ml to 833.33?8.73 ii g/ml, the expression ratio of mIL-4 on CD4 CDs cell membrane rises from 6.8 .2% to 29.3 .4%(p |
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