Transplant Human PBMC To Establish A "Human-mouse" Xenogeneic Graft-versus-host Disease Model

Experiments in the human body are strictly limited as moral, ethical, legal and other aspects of the constraints. Research involving human cells generally can only be done in vitro, but the results usually have some differences. Establishment of "Human - mouse" model provides the possibility of research in vivo. However, there is a lot of reports about the "Human-mouse" model, but little of them is effective and stable. "Human-mouse" model is most widely used for xenogeneic graft versus host disease model, but the methods also have certain differences. Our work is selecting the appropriate strains of mice and optimizing of experimental conditions to establish a stable"Human-mouse"xenogeneic graft-versus-host disease model for inducing T cell tolerance research. The following is a brief introduction to this research:This study selects the Nude Mice and NOD / SCID, and gives them sublethal dose ofγ-ray irradiation whole body, and then intraperitoneal transplant human peripheral blood mononuclear cells (PBMC) to establish xenogeneic acute graft-versus-host disease model. By detection of human T cells in the mice's tail venous blood, tissues, organs of the infiltration and other indicators (By flow cytometry and immunohistochemistry), to compare the human immune cell infiltration rates in the two mouse model and record the survival time. Finally, determining the appropriate strains of mice to establish X-GVHD. Optimization means are the transfer ways and the appropriate amount of human PBMC, and the best time to observe the changes of T cell phenotype and function.The results show that the NOD/SCID mouse is more suitable for inducing"Human-mouse"X-GVHD model, and there are no significant differences between intraperitoneal injection and intravenous injection. Transfer human PBMC more then 5×10~7 can establish"Human-mouse"X-GVHD model. Use the optimized experimental conditions to establish the"Human-mouse"X-GVHD model, we find the best time to observe the changes of T cell phenotype and function is between 7-11 days, and the average survival time is 14.16±1.77 days.