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【目的】为了探索治疗急性乙型肝炎和爆发性乙型肝的新策略,本研究将HBV DNA疫苗和相应抗原的蛋白质分子联合免疫小鼠,旨在探讨联合免疫对小鼠抗原特异性T细胞增殖反应的影响。【方法】我们将HBV DNA疫苗(pcDS2)和相应抗原蛋白质分子(HBsAg)联合免疫BALB/c小鼠;分别在第0、2和4周进行免疫,在第6周用ELISA方法检测抗-HBs IgG效价,MTT和流式细胞仪检测T细胞增殖反应及流式细胞仪检测细胞因子表达水平。【结果】pcDS2和HBsAg联合免疫组小鼠的抗-HBs水平显著提高;免疫小鼠的T细胞体外经HBsAg刺激后,联合免疫组刺激指数(SI)明显降低;经流式细胞仪检测进一步证实联合免疫组T细胞增殖反应被显著抑制;联合免疫组T细胞表达IL-10和Foxp3水平显著升高。【结论】pcDS2和HBsAg联合免疫能诱导产生特异性体液免疫应答,但不能诱导产生抗原特异性T细胞增殖反应;T细胞增殖反应被显著抑制可能与T细胞表达IL-10和Foxp3上调有关;本研究为急性乙型肝炎和爆发性乙型肝炎治疗及HBV疫苗的研究奠定了基础。
【Objective】 In order to explore a new strategy for the treatment of acute hepatitis B and fulminant hepatitis B, we combined immunization with HBV DNA vaccine and protein molecules of corresponding antigens to explore the effect of combined immunization on mouse antigen-specific T cells Effect of proliferative response. 【Methods】 BALB / c mice were immunized with HBV DNA vaccine (pcDS2) and corresponding antigenic protein (HBsAg). Immunization was performed at weeks 0, 2 and 4, and anti-HBs IgG titer, MTT and flow cytometry to detect T cell proliferative response and flow cytometry cytokine expression levels. 【Results】 The results showed that the anti-HBs levels in the mice immunized with pcDS2 and HBsAg were significantly increased. The T cells of the immunized mice were significantly decreased after stimulation with HBsAg in vitro. The results of flow cytometry T cell proliferation response was significantly inhibited in the co-immunized group; the levels of IL-10 and Foxp3 in the T cells in the co-immunized group were significantly increased. 【Conclusion】 Combined immunization with pcDS2 and HBsAg can induce specific humoral immune response, but can not induce antigen-specific T cell proliferative response; T cell proliferation was significantly inhibited may be related to T-cell upregulation of IL-10 and Foxp3; The research lays the foundation for the research on the treatment of acute hepatitis B and hepatitis B and HBV vaccine.