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目的观察日本血吸虫感染小鼠调节性T细胞水平动态变化,筛选调节性T细胞功能抑制因子。方法以日本血吸虫尾蚴感染小鼠,采用流式细胞分析法检测不同感染阶段小鼠体内调节性T细胞水平;通过检测小鼠血清中细胞因子的水平变化,观察免疫反应的极化过程;采用磁珠分离法纯化小鼠脾脏的调节性T细胞,人工设计合成2条寡核苷酸(CpG1-ODN,CpG2-ODN),通过淋巴细胞增殖试验筛选能逆转调节性T细胞功能的刺激因子。结果流式细胞分析发现,随着血吸虫感染进程的发展,小鼠外周血及脾脏调节性T细胞水平逐渐增加,免疫反应逐步向Th2型极化,血清IL-5水平增加,IFN-γ下降。淋巴细胞增殖试验显示,人工合成的寡核苷酸(CpG2-ODN)能有效刺激淋巴细胞增殖,使淋巴细胞的IL-2分泌增加,TGF-γ下降,并能解除调节性T细胞对效应T细胞增殖的免疫抑制作用。结论血吸虫感染可增加小鼠体内调节性T细胞水平,诱导免疫反应向Th2型极化。寡聚核苷酸CpG2-ODN能逆转调节性T细胞的免疫抑制功能。
Objective To observe the dynamic changes of regulatory T cells in mice infected with Schistosoma japonicum and screen the regulatory T cell function inhibitors. Methods Mice infected with Schistosoma japonicum cercariae were used to detect the level of regulatory T cells in mice during different stages of infection by flow cytometry. The level of cytokines in serum was detected to observe the polarization of immune response. Purification of regulatory T cells in spleen of mice by bead separation and two oligonucleotides (CpG1-ODN, CpG2-ODN) were artificially designed and synthesized. Stimulation factors capable of reversing the function of regulatory T cells were screened by lymphocyte proliferation assay. Results The results of flow cytometry showed that with the progression of schistosomiasis infection, the level of T regulatory cells in peripheral blood and spleen of mice gradually increased, the immune response gradually increased to Th2 type, the level of serum IL-5 and the level of IFN-γ decreased. Lymphocyte proliferation test showed that the synthetic oligonucleotide (CpG2-ODN) can effectively stimulate lymphocyte proliferation, lymphocyte IL-2 secretion, decreased TGF-γ, and can relieve regulatory T cells on the effect of T Immunosuppressive effects of cell proliferation. Conclusion Schistosoma infection can increase the level of regulatory T cells in mice and induce the immune response to Th2-type polarization. Oligonucleotide CpG2-ODN can reverse the immunosuppressive function of regulatory T cells.