CpG-ODN增强乙型肝炎病毒转基因小鼠对重组乙肝疫苗的细胞免疫应答

CpG-ODN potently enhances cellular immune responses in HBV transgenic mice induced by recombinant hepatitis B vaccine

关键词:  CpG-ODN  重组酵母乙肝疫苗  乙肝病毒转基因小鼠  细胞免疫应答

Key words:  CpG oligodeoxynucleotides  recombinant hepatitis B vaccine  HBV Transgenic Mice  cellular immune responses

摘要:

[目的 ]探讨CpG -ODN作为佐剂对重组乙肝疫苗诱导乙肝病毒转基因小鼠产生细胞免疫应答的影响。[方法 ]用重组干扰素 (rIFNα - 2b)、重组乙肝疫苗 (rHBsvaccine)和重组乙肝疫苗 +CpG -ODN分别免疫HBV转基因小鼠后 ,流式细胞仪检测免疫小鼠的外周血T淋巴细胞亚群 ,ELISA法检测免疫小鼠的外周血IL - 2、IL - 12 (p70 )和IFN -γ的含量 ,MTT法和乳酸脱氢酶法分别检测免疫小鼠的淋巴细胞增殖和杀伤功能。[结果 ]重组乙肝疫苗组CD3+、CD4 +、CD8+细胞在T细胞中所占百分比分别为 (2 5 .0 0 8± 2 .85 2 ) %、(13.6 40± 1.6 16 ) %、(9.989± 1.12 3) %均明显高于对照组 ,(P <0 .0 5 ) ;IL - 2、IL - 12 (p70 )和IFN -γ的含量分别为 (130 4 .2 88± 5 72 .2 12 ) pg/mL、(2 0 3.810± 91.80 7) pg/mL、(86 0 .736± 336 .888)pg/mL均明显高于对照组 ,(P <0 .0 5 ) ;淋巴细胞特异性增殖和杀伤效应与对照组相比均有明显差异 ,(P <0 .0 5 )。疫苗 +CpG -ODN组与疫苗组比较 ,免疫小鼠产生更强的HBV特异性细胞应答 (P <0 .0 5 ) ,且以Th1型细胞免疫应答为主。而干扰素组由于给药次数和剂量等原因 ,与对照组无显著差异。 [结论 ]CpG -ODN作为佐剂可以较好的增强重组乙肝疫苗诱导HBV转基因小鼠产生细胞免疫应答。

Abstract:

[Objective] To investigate the effects of CpG-ODN as adjuvants on cellular immune responses in HBV Transgenic Mice immunized with recombinant hepatitis B vaccine. [Methods] After HBV transgenic mice were immunized with rIFNα-2b、recombinant hepatitis B vaccine and recombinant hepatitis B vaccine + CpG-ODN, the subsets of T lymphocyte were investigated through FACSVantageTMSE, IL-2、IL-12(p70)and IFN-γ were examined by ELISA, the proliferation of spleen cells was measured by MTT and cytotoxic T lymphocyte (CTL) response was tested by lactic acid dehydrogenase (LDH) releasing method. [Results] CD3+%、CD4 +%、CD8 +% of recombinant hepatitis B vaccine group were(25.008±2.852)%、(13.640±1.616)%、 (9.989 ±1.123)%; the amount of IL-2、IL-12(p70)、IFN-γwere (1304.288±572.212)pg/mL、 (203.810 ±91.807)pg/mL、(860.736±336.888)pg/mL . Compared with N.S., recombinant hepatitis B vaccine could distinctly raise CD3 +%、CD4 +%、CD8 +% and the amount of IL-2、IL-12(p70)、IFN-γ, and induce strong lymphocyte proliferation reaction and cytotoxic T lymphocyte activities (P<0.05). Immunized HBV Transgenic Mice with CpG-ODN showed higher specific cellular responses to HBV than those without CpG-ODN (P<0.05), and the responses dominantly were Th1 cellular immune responses. Because of times and doses of rIFNα-2b, immunized with rIFNα-2b and N.S. have no obvious difference. [Conclusion] CpG-ODN can act as adjuvants to enhance cellular immune responses in HBV Transgenic Mice immunized with recombinant hepatitis B vaccine.