中华皮肤科杂志

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白介素2基因佐剂协同HSV-2多表位复合DNA疫苗诱导小鼠免疫应答

徐金华1, 许炎2, 郑志忠1, 夏天2, 王健1, 梁俊1, 张臻1, 朱小华1, 朱乃硕2   

  1. 1. 复旦大学附属华山医院皮肤科, 上海, 200040;
    2. 复旦大学生命科学学院病毒与分子免疫学研究室
  • 收稿日期:2006-10-23 出版日期:2007-07-15 发布日期:2007-07-15
  • 通讯作者: 朱乃硕,email:Zhunaishuo@sina.com E-mail:Zhunaishuo@sina.com
  • 基金资助:
    上海市科委资助课题(024119011)

Induction of immune response in mice by combined immunization with HSV-2 multi-epitope DNA vaccines and interleukin-2 genetic adjuvant

XU Jin-hua1, XU Yan2, ZHENG Zhi-zhong1, XIA Tian2, WANG Jian1, LIANG Jun1, ZHANG Zhen1, ZHU Xiao-hua1, ZHU Nai-shuo2   

  1. Department of Dermatology, Huashan Hospital, Fudan University, Shanghai 200040, China
  • Received:2006-10-23 Online:2007-07-15 Published:2007-07-15

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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ICP27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±219.60ng/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-2cDNA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.

关键词: 疱疹病毒2型, 人, 单纯疱疹病毒疫苗, 白细胞介素2, 糖蛋白类, 感染细胞蛋白27

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 HVpcDNA3.1,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 combination 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 HVpeDNA3.1 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.

Key words: Herpes virus 2,human, Herpes simplex virus vaccines, Interleukin-2, Glycoproteins, Infectious cell protein 27