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目的 观察编码鼠IL-12的真核表达载体对HBV DNA疫苗诱导Balb/c小鼠免疫应答的影响。 方法 肌肉注射DNA疫苗,ELISA法检测小鼠血清抗HBs,4h ~(51)Cr释放法检测小鼠脾细胞CTL杀伤活性。 结果 免疫8wk后,注射pCR3.1组、注射pCR3.1-S组及共注射IL-12真核表达载体的血清A(OD值)分别为0.04±0.01,0.87±0.1及1.67±0.15。CTL细胞杀伤活性分别为10.1％±6.4％,50.5％±6.4％及73.3％±8.8％,后两组与pCR3.1组比较均有显著差异(P<0.01),后两组比较均有显著差异(P<0.01).脾细胞悬液经抗CD4~+单克隆抗体处理后分别为48.3％±5.9％,75.6％±9.1％,抗CD8~+单克隆抗体处理后分别为10.6％±1.4％,16.9％±2.3％。 结论 IL-12的真核表达载体能够提高小鼠对DNA疫苗的免疫应答,CTL细胞杀伤活性主要由CD8~+执行。基因疫苗可能用于预防及治疗HBV感染。
Objective To observe the effect of eukaryotic expression vector encoding murine IL-12 on immune response induced by HBV DNA vaccine in Balb / c mice. Methods DNA vaccines were injected intramuscularly. Serum anti-HBs were detected by ELISA. CTL killing activity of spleen cells was detected by 4h ~ (51) Cr release assay. Results After 8 weeks of immunization, serum A (OD) of pCR3.1 injection group, pCR3.1-S injection group and co-injection of IL-12 eukaryotic expression vector were 0.04 ± 0.01, 0.87 ± 0.1 and 1.67 ± 0.15, respectively. CTL cell killing activity was 10.1% ± 6.4%, 50.5% ± 6.4% and 73.3% ± 8.8%, respectively. There was a significant difference between the latter two groups and the pCR3.1 group (P <0.01) (P <0.01) .The splenocytes suspension were treated by anti-CD4 ~ + monoclonal antibody, they were 48.3% ± 5.9% and 75.6% ± 9.1% respectively after treated with anti-CD4 ~ + monoclonal antibody, and 10.6% ± 1.4 %, 16.9% ± 2.3%. Conclusion The eukaryotic expression vector of IL-12 can improve the immune response of mice to DNA vaccine. The killing activity of CTL is mainly performed by CD8 +. Gene vaccines may be used to prevent and treat HBV infection.