丙型肝炎病毒重组联合疫苗的实验研究

Experimental study of a HCV recombinant combined vaccine

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 HCV recombinant combined vaccine based on several antigens of HCV. Methods：Mice were co-immunized with three recombinant antigens F4HVR1, HCV-T and HCV-E1. The antibodies in sera were tested by ELISA. Spleens from immunized BALB/c mice were removed 10 days after the third immunization. CTL activity was assessed using a LDH Cytotoxicity Assay Kit. IFN-γ- and IL-4- secreting cells were quantified using a ELISPOT kit. Percentages of CD4^＋ T and CD8^＋ T cells in the spleen cells of mice were analyzed with flow cytometer. Two weeks after the final immunization, the mice were challenged s. c. at the back with 1.0 × 10^6 SP2/0-NS3 cells, and protective effect was obaerved. Results：Co-immunization with F4HVR1 ＋ HCV-T＋ HCV-E1 induced high tiers of HCV-E1- or F4HVR1- specific IgG antibody,high level of CrL activity,and a balanced Th1/Th2 response. The combined vaccine F4HVRI ＋ HCV-T ＋ HCV-E1 could significantly protect mice from SP2/O-NS3 cells. ： Co-immunization with F4HVR1 ＋ HCV-T ＋ HCV-E1 induces protective humoral and cellular immune responses.F4HVR1 ＋ HCV-T ＋ HCV-E1 is potential as a prophylactic HCV vaccine.