The Influence Of Foxp3 Gene-transferred CD4+CD25- T Cells On The Survival Of Heterotopic Heart Transplantation In Mice

CD4+CD25+ regulatory T cell has attracted wide attention in the field of organ transplantation, for its ability of immune suppression, and is considered as one of effective methods in inducing immune tolerance after organ transplantation. But so far there isn't any report about its clinical application, the most important reason may be it's quantity limitation. The effort of expanding them in vitro on the condition in vivo has brought some results, But this don't take any advantage to clinical work, for it need a long time culture, the high cost, and the severe experimental condition. The identify of Foxp3 provides us another pathway to obtain this T cell subset. Foxp3 gene-transferred CD4+CD25-T cell has the affinis cell phenotype and the ability of immune suppression with the nature occurring CD4+CD25+ regulatory T cell. This make it nearer realization to harvest enough regulatory T cells.Objective:To transfer Foxp3 gene to mice CD4+CD25-T cell via retroviral Vector pLXSN, and make it obtain the ability of immune suppression. To study the ability of these cells suppressing lymphocyte proliferation in vitro, and it's influence on the the survival of heterotopic heart transplantation in miceMethod:1. First of all the retroviral system of Foxp3, pLXSN-Foxp3-IRES2-EGFP, will be constructed via the primitive plasmid pMD18-T-Foxp3. Then produce virus supernatant through package cells PA317, and infect CD4+CD25-T lymphocyte isolated form spleen of C57BL/6 mice, inlet Foxp3 gene and report gene EGFP into target cells, Appraise the transfection efficiency of this retroviral system through observing the expressing of EGFP and the detection of flow cytometry. 2. CD4+CD25+ regulatory T cells and Foxp3 gene-transferred CD4+CD25-T cells will...