小鼠对HBV DNA疫苗的免疫应答及细胞因子的免疫佐剂效应  

The immune responses to hepatitis B gene vaccine in mice and the immune adjuvant effect of cytokines

作　　者：杜德伟[1] 周永兴[1] 白宪光[1] 冯志华 李光玉[1] 姚志强[1] 

机构地区：[1]第四军医大学唐都医院感染防治中心,陕西西安710038

出　　处：《医学研究生学报》2001年第2期95-99,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金!资助项目 (编号 :3 9770 665 )

摘　　要：目的 :观察编码小鼠 IL- 2及 IL- 12的真核表达载体 ,对 HBV DNA疫苗诱导 Balb/ c小鼠 (H- 2 d )免疫应答的调节作用及其对稳定表达 HBs Ag的小鼠肥大细胞瘤 P815细胞 (P 815 - HBV- S)成瘤性的影响。　方法 :肌内注射HBV DNA疫苗及细胞因子真核表达载体 ;背部皮下接种 P 815 - HBV - S细胞 ,观察成瘤情况 ;EL ISA法测定血清抗HBs;4h5 1 Cr释放法检测小鼠脾细胞 CTL 活性。　结果 :免疫 8周后 ,以 p CR3.1- S、p CR3.1- S+IL- 2及 p CR3.1-S+IL- 12免疫的小鼠血清 ,45 0 nm A值分别为 0 .87± 0 .1、1.98± 0 .17及 1.6 7± 0 .15 ,均较 p CR3.1组显著提高(P<0 .0 5 )。CTL 细胞杀伤活性分别为 (5 0 .5± 6 .4) %、(6 1.9± 7.1) %及 (73.3± 8.8) % ,后两组与 p CR3.1- S组比较差异均显著 (P<0 .0 5 )。脾细胞悬液经抗 CD4+ 单克隆抗体处理后 ,CTL细胞杀伤活性分别为 (48.3± 5 .9) %、(5 6 .2± 7.5 ) %和 (75 .6± 9.1) % ,抗 CD8+ 单克隆抗体处理后分别为 (10 .6± 1.4) %、(13.6± 1.9) %和 (16 .9±2 .3) %。对照组成瘤率为 10 0 % ,p CR3.1- S组小鼠成瘤率为 12 .5 % ,p CR3.1- S+IL- 12真核表达载体组成瘤率为0 ,对照组平均存活期和 6周后存活率分别为 2 8.4天和 0天。后两组分别 >38.2天及 45?Objectives:To observe the effect of eukaryotic expression vectors coding IL 2 and IL 12 on immune responses induced by DNA immunization of HBV surface antigen(pCR3.1 S)in BABL/c(H 2 d) and the protection against P815 mastocytoma cells stable expressing HBV surface antigen in mice after immunized with HBV gene vaccine. Methods:The immunization was performed by intramuscular injection, three weeks later, we directly inoculated P815 HBV S into mice by subcutaneous injection .Tumor growth was measured every five days. Anti HBs in serum was detected by ELISA and HBsAg specific cytotoxic T lymphocytes (CTLs) activity was measured by 51 Chromium release assay. Results:Eight weeks after immunization, the A value of mice serum in 450 nm and CTLs activity of mice codiog IL 2 and IL 12 eukaryotic expression vectors were significant higher( P <0.05) than that of mice intramuscular injected HBV S DNA vaccine, these values are significant higher than that of mice injected pCR3.1( P <0.05). The spleen cells CTLs activity have decreased obviously after treated with anti CD8 + monoclonal antibody and have no significant change after treated with anti CD4 + monoclonal antibody. The HBV S gene vaccine could evidently inhibit the tumor growth, prolong the survival period (>38.2 days) and improve the survival rate in mice. Conclusions:The DNA vaccine of HBV ( pCR3.1 S) had strong antigenicity in cellular and humoral immunity and had marked killing effect on HBV infected cells in vivo ,which could be promoted by vector coding murine IL 2 or IL 12. CTLs activity was performed by CD8 + cells.

关 键 词：DNA疫苗 乙型肝炎表面抗原 细胞因子 小鼠 真核表达载体 

分 类 号：R446.62[医药卫生—诊断学]