1. The Study Of NOD.H-2～(h4) Mice T Cell Cloning 2. The Constructs Of HPV, HSV Recombinated Prophylactic Vaccine

ObjectiveA case in point is experimental autoimmune thyroiditis, which is the example of autoimmune diseases. The ability to generate, maintain, and use cloned lines of T cells reactive for self-antigens has opened up a new avenue of investigation for researchers. To elucidate the immune function of T cell and B cell and T cell cloning, which were stimulated by Tg (thyroglobulin, Tg) presenting dendritic cells (DCs) in experimental autoimmune thyroiditis from bone marrow of NOD.H-2 h4 mice and to elucidate the mechanism of induction in experimental autoimmune thyroiditis(EAT), Antigen specific T cell clones have been investigated by dendritic cell as the antigen-presenting cells primed with thyroglobulin(Tg).Materials and MethodsNOD.H-2 h4 (Non-obese-Diabetes) mice under SPF(specific-pathogen free) condition were given water with 0.05% iodine for 8 weeks and mice were bled biweekly intervals to measure their level of Tg by ELISA then some of them were immunized Tg 100ug per mouse in complete Freund adjuvant twice. Before sacrificed those immunized mice, proliferation assay has been done to optimize the amount of Tg for the dendritic cells to uptake in order the antigen-presenting cells to present Tg to T cells. Finally, splenocytes taken from Tg immunized mice were co-cultured with dendritic cells from bone marrow of syngeneic mice, which were cultured, irradiated and incubated with Tg as the feeder cells. We also did passive transfer experiment to NOD.H-2 h4 mice by using T cell clones. That is, four out of six T cell clones, F11, 2D11, H8, G8, were transferred to either group of normal NOD.H-2 h4 mice or group of irradiated NOD.H-2 h4 mice.ResultsSix Tg-specific T cell clones were established and they were cultured in vitro over two years and all of them proliferated to Tg and their phenotype are CD4+ by FACS analysis as well as secreted IFN-γinstead of IL-4.Passive transfer experiment by T cell clones indicated four T cell clones were not caused thyroiditis in the normal NOD.H-2 h4 mice and three out of four clones can induce thyroiditis in irradiated NOD.H-2 h4 mice and irradiated dose (475 RAD) to the NOD.H-2 h4 mice is sufficient to remove the function of suppressive cells in receipt mice those have the function to help B cell to produce antibody.ConclusionsThe ability to generate, maintain, and use cloned lines of T cells reactive for self-antigens has opened up a new avenue of investigation for researchers. Dendritic cells, which act as Tg...