利用体内电脉冲增强治疗型HBV DNA疫苗细胞免疫反应研究.docVIP

利用体内电脉冲增强治疗型HBV DNA疫苗细胞免疫反应研究 　　[摘要] 目的:研究在初次免疫和加强免疫阶段分别使用不同的免疫模式能否增强HBV DNA疫苗的细胞免疫反应。方法:肌内注射10 μg HBV DNA疫苗的Balb/c小鼠根据电脉冲实施时间的不同选择两种免疫模式,DNA+EP或EP+3 d+DNA,按在初次免疫和加强免疫阶段免疫模式的不同组合次序进行分组。各组小鼠在实施加强免疫后第14天取脾细胞做抗原特异的IFN-γ ELISPOT检测和FACS分析。结果:在初次免疫和加强免疫阶段使用不同免疫模式的实验组与采用相同免疫模式的实验组相比,诱导的特异性IFN-γ T细胞频数和IFN-γ的CD8+ T细胞百分比均增加了4倍以上,差异有高度统计学意义(P0.05)。结论:在初次免疫和加强免疫阶段利用体内电脉冲实施不同的免疫模式能显著增强HBV DNA疫苗在动物体内的细胞免疫效果。 　　[关键词] HBV DNA疫苗;体内电脉冲;初次免疫-加强免疫策略;细胞免疫反应 　　[中图分类号] R392[文献标识码]A [文章编号]1673-7210(2010)04(b)-023-04 　　 　　Alternating immune modes utilizing electric pulses in vivo enhance cellular immune responses induced by therapeutic HBV DNA vaccine 　　QIU Ligong, SUN Xihai, WEI Jian, ZHAO Liang, HUANG Ming 　　(Baidi Bio-Pharmaceutical RD Center, Guangzhou Pharmaceutical Holdings Limited, Guangzhou 511495, China) 　　[Abstract] Objective: To investigate the possibility of improving cellular immune responses induced by HBV DNA vaccine by alternating immune modes in prime-boost regimen. Methods: Balb/c mice to be vaccinated with 10 μg HBV DNA vaccine were divided according to the order of immune modes which could be chosen between two alternative immune modes (DNA+EP or EP+3 d+DNA) and could be same or alternating in prime-boost regimen. The splenocytes from mice of all groups were detected by antigen-specific IFN-γ ELISPOT and FACS assays at the 14 th day after boost. Results: The numbers of antigen-specific IFN-γ secreting T cell or CD8+ T percentage from groups of alternating immune modes were 4 times higher than those of the same immune modes (P0.05). Conclusion: Alternating immune modes in prime-boost regimen utilizing electric pulses can induce high levels of cellular immune responses. 　　[Key words] HBV DNA vaccine; Electric pulses in vivo; Prime-boost strategy; Cellular immune responses 　　 　　近年来,以质粒DNA为基础???免疫方法广泛应用于疾病预防和治疗研究中,显示了巨大的治疗潜力。为了提高质粒DNA的免疫原性和免疫效果,学者们进行了各种方法和策略的研究,其中以电脉冲(electric pulse,EP)介导的DNA疫苗免疫方法在激发高水平免疫反应方面具有突出的效果。EP能增强DNA疫苗的免疫应答,其效应不受实验动物种属的限制[1-2]。现有的研究和临床方案