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【目的】为了进一步增强HBV DNA疫苗的免疫反应,本研究将共刺激分子OX40L作为HBV DNA疫苗的分子佐剂免疫小鼠,旨在探讨共刺激分子OX40L对HBV DNA疫苗诱导体液和细胞免疫应答的影响。【方法】我们将HBV DNA疫苗(pcDS2)单独或联合共刺激分子质粒pOX40L免疫C57BL/6小鼠;分别在第0,2,4周进行免疫,在第6周检测抗-HBsIgG、IgG1和IgG2a,T淋巴细胞增殖指数,细胞因子表达水平和体内细胞毒性T淋巴细胞杀伤作用(CTL)等免疫学指标。【结果】pcDS2联合pOX40L免疫组小鼠的抗-HBs水平显著提高,抗-HBsIgG亚类以IgG2a占优;免疫小鼠的T淋巴细胞体外经乙型肝炎表面抗原(HBsAg)刺激后,联合免疫组刺激指数(SI)明显高于pcDS2组;联合免疫组CD4+T淋巴细胞的IL-4和IFN-γ表达水平及CD8+T淋巴细胞的IFN-γ表达水平显著升高;DNA疫苗免疫的各组小鼠,HBsAg特异性体内CTL高于对照组,其中联合免疫组小鼠的体内CTL杀伤作用最强。【结论】共刺激分子OX40L不仅能增强HBV DNA疫苗诱导特异性体液免疫应答,还能增强特异性细胞免疫反应,尤其增强体内CTL的杀伤活性,为HBV DNA疫苗的研究奠定了基础。
【Objective】 In order to further enhance the immune response of HBV DNA vaccine, this study used costimulatory molecule OX40L as a molecular adjuvant to HBV DNA vaccine to immunize mice to explore the effect of costimulatory molecule OX40L on humoral and cellular immune responses induced by HBV DNA vaccine influences. 【Methods】 C57BL / 6 mice were immunized with pcDNA2 (pcDS2) alone or in combination with costimulatory plasmid pOX40L. Immunized at weeks 0, 2 and 4, and anti-HBs IgG, IgG1 and IgG2a , T lymphocyte proliferation index, cytokine expression and cytotoxic T lymphocyte cytotoxicity (CTL) and other immunological indicators. [Results] The anti-HBs levels of pcDS2 combined with pOX40L immunized mice were significantly increased, and the anti-HBs IgG subgroup was dominated by IgG2a. The T lymphocytes of immunized mice were stimulated with HBsAg in vitro, (SI) was significantly higher than that of pcDS2 group. The expression of IL-4 and IFN-γ in CD4 + T lymphocytes and the expression of IFN-γ in CD8 + T lymphocytes in combination group were significantly increased. Each group of mice, HBsAg-specific in vivo CTL higher than the control group, of which the combined immunization group mice CTL strongest killing effect in vivo. 【Conclusion】 Costimulatory molecule OX40L can not only enhance the specific humoral immune response induced by HBV DNA vaccine but also enhance the specific cellular immune response, especially CTL cytotoxic activity in vivo, which lays the foundation for the study of HBV DNA vaccine.