【摘要】 目的研究白介素2(IL-2)cDNA协同HSV-2多表位复合DNA疫苗免疫对小鼠体液和细胞免疫应答的影响。方法利用真核表达质粒pcDNA3.1分别构建HSV-2糖蛋白gD、gB、gC和早期表达蛋白ICP 27的多表位复合DNA疫苗HV-pcDNA3.1和IL-2的真核表达质粒IL-2-pcDNA3.1。15只C57/ BL6小鼠分为3组,分别接受pcDNA3.1空载体注射、HV-pcDNA3.1单独免疫或HV-pcDNA3.1及IL-2- pcDNA3.1联合免疫。共免疫2次,间隔2周。末次免疫3周后。ELISA法检测小鼠血清HSV-2特异性IgG、IL-2和IFN-γ,淋巴细胞特异性增殖反应和乳酸脱氢酶法检测杀伤性T淋巴细胞(CTL)功能。结果与HV-pcDNA3.1单独免疫组相比.IL-2-pcDNA3.1的协同免疫可以提高小鼠血清中和IFN-γ表达(1956.19±219.60 ng/L,P<0.05),增强淋巴细胞特异性增殖反应(促细胞增殖率为2.75%±0.26%,P<0.05)和CTL活性(47.91%±15.09%,P<0.05)。小鼠血清HSV-2特异性IL-2水平在协同免疫组为(1421.16±220.98)ng/L,HV-pcDNA3.1单独免疫组为(1246.84±157.02)ng/L.两组间差异无统计学意义。结论IL-2 cDNA的协同免疫可以显著提高HSV-2多表位复合DNA疫苗诱导的细胞免疫应答水平。更多还原

【Abstract】 Objective To investigate the effect of combined immunization of interleukin-2(IL-2) eDNA and HSV-2 muiti-epitope DNA vaccines on the humeral and cellular immunity of mice.Methods The eukaryotic expression plasmid pcDNA3.1 was used to construct a multi-epitope DNA vaccine HV- pcDNA3.1,which contained the genes of glycoprotein D,glycoprotein B,glycoprotein C and infectious cell protein 27(ICP 27),and a recombinant eukaryotic expression plasmid IL-2-pcDNA3.1.Fifteen C57/BL6 mice were divided into 3 groups immunized with pcDNA3.1 vector,HV-pcDNA3.1 vaccine,and combina- tion of HV-pcDNA3.1 vaccine and IL-2-pcDNA3.1,respectively for 2 times with an interval of 2 weeks. Three weeks after the last immunization,ELISA was used to detect the serum levels of HSV-specifie IgG, IL-2 and IFN-γin these mice,lactate dehydrogenase assay to evaluate the function of CTL,and MTT method to assess the lymphproliferation response of T lymphocytes collected from the spleen of these mice. Results In comparison with the immunization with HV-pcDNA3.1 vaccine alone,the coinjection of IL-2-pcDNA3.1 and HV-pcDNA3.1 increased the serum levels of IFN-γ(1956.19±219.60 vs 1340.27±108.52 ng/L,P<0.05),and CTL activity(47.91%±15.09% vs 31.82%±10.12%,P<0.05).Also,the combined immunization elevated the proliferation rate of T lymphocytes more significantly than HV- peDNA3.1 vaccine alone(2.75%±0.26% vs 1.18%±0.06%,P<0.05).The serum levels of specific IL-2 were 1246.84±157.02 ng/L and 1421.16±220.98 ng/L in mice immunized by HV-pcDNA3.1 alone and those by IL-2-pcDNA3.1 and HV-pcDNA3.1,respectively,but no statistical difference was found between them.Conclusion IL-2 eDNA may enhance the celluar immune response induced by HSV-2 multi-epitope DNA vaccine in mice.更多还原