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目的:研究一种基于丙型肝炎病毒(HCV)多个抗原的重组联合疫苗。方法:将三种HCV重组抗原F4HVR1、HCV-T和HCV-E1混合,联合免疫BALB/c小鼠,用ELISA检测血清抗体滴度;第三次免疫后10天杀死一半小鼠,取脾细胞,用乳酸脱氢酶(LDH)释放法检测细胞毒性T淋巴细胞(CTL)活性;用ELISPOT法检测分泌IFN-γ和IL-4的细胞;用流式细胞仪检测CD4+T和CD8+T细胞;最后一次免疫后2周,在免疫小鼠的背部皮下注射1.0×106个SP2/0-NS3细胞,考察联合免疫对小鼠的保护作用。结果:F4HVR1+HCV-T+HCV-E1联合免疫诱导了高滴度的HCV-E1和F4HVR1特异性的IgG抗体以及高水平的CTL活性;与单独免疫相比,联合免疫诱导了平衡的Th1/Th2免疫应答;而且联合免疫后能显著预防SP2/0-NS3对小鼠的攻击。结论:F4HVR1+HCV-T+HCV-E1诱导了保护性的体液免疫和细胞免疫应答,有望开发为一种有效的重组HCV联合疫苗。
Objective: To study a recombinant combined vaccine based on multiple antigens of Hepatitis C virus (HCV). Methods: BALB / c mice were immunized with three HCV recombinant antigens F4HVR1, HCV-T and HCV-E1, and serum antibody titers were detected by ELISA. Half of the mice were killed 10 days after the third immunization, The cytotoxic T lymphocyte (CTL) activity was detected by lactate dehydrogenase (LDH) release assay. The cells secreting IFN-γ and IL-4 were detected by ELISPOT assay. The levels of CD4 + T and CD8 + T cells. Two weeks after the last immunization, 1.0 × 106 SP2 / 0-NS3 cells were subcutaneously injected into the back of the immunized mice to investigate the protective effect of combined immunization on mice. Results: Co-immunization with F4HVR1 + HCV-T + HCV-E1 induced high titers of HCV-E1 and F4HVR1-specific IgG antibodies as well as high levels of CTL activity; co-immunization induced a balanced Th1 / Th2 immune response; and combined immunization can significantly prevent the SP2 / 0-NS3 attack on mice. CONCLUSION: F4HVR1 + HCV-T + HCV-E1 induces protective humoral and cellular immune responses and is expected to be developed as a potent recombinant HCV combined vaccine.