治疗性HBV DNA疫苗诱导健康Balb/c小鼠细胞免疫应答的实验研究  

Cellular immune responses induced by therapeutic HBV DNA vaccine in healthy Balb/c mice

作　　者：刘惠萍[1] 杨富强[1] 饶桂容 莫国玉[1] 陈光明[1] 

机构地区：[1]解放军第458医院全军肝病中心,广州510602

出　　处：《免疫学杂志》2006年第6期612-614,共3页Immunological Journal

基　　金：国家"863"计划资助项目(2001AA217141)

摘　　要：目的探讨治疗性HBVDNA疫苗对健康Balb/c小鼠表达Th1类细胞免疫应答的影响。方法治疗性HBVDNA疫苗(pS2·S+pFP)以剂量(50μg+50μg)/只对Balb/c小鼠进行接种,以酶联免疫斑点法(ELISPOT)、流式细胞术(FACS)分析特异性分泌IFN-γ阳性T细胞的免疫效果。结果免疫6周时,实验组用HBsAg刺激后其诱生的IFN-γ细胞频数(34±15.3)较不用HBsAg组(4±4.4)高(P<0.05,t=5.65),也较同期空载体免疫对照组(3±3.6)高(P<0.05,t=5.21);治疗性HBVDNA疫苗诱导小鼠CD4+T细胞表达IFN-γ的百分比实验组[(0.28±0.17)%]与对照组[(0.20±0.03)%]比较,无显著性差异(P>0.05,t=0.21);而诱导CD8+T细胞表达IFN-γ的百分比实验组[(0.69±0.08)%]与对照组[(0.24±0.12)%]比较,具显著性差异(P<0.05,t=4.167)。结论治疗性HBVDNA疫苗能有效诱导T细胞分泌HBsAg特异性IFN-γ因子,并以CD8+T细胞分泌占优而发挥细胞免疫应答的优势。Objective To investigate the cellular immune responses induced by HBV DNA vaccine in healthy Balb/c mice. Methods HBV DNA vaccine （ pS2- S ＋ pFP） was vaccinated in healthy Balb/c mice with the dosage of （50μg ＋ 50μg） per mouse, and then the immune responses of IFN-γ-secreting cells （SFC/10s splenocytes） and CD4^＋/CD8^＋ T lymphocytes after immunization were detected by ELIS- POT and FACS assays. Results At the 6^th week after immunization of HBV DNA vaccine, the level of IFN-γ in HBsAg-stimulated group （34 ± 15.3） was much higher than that of unstimulated group in vitro （4±4.4） （ P 〈 0.05, t = 5.65） and that of control group （3 ±3.6） （ P 〈 0.05, t = 5.21 ）. The percentage of IFN-γ-secreting CD4^＋ T cells in the experimental group [ （0.28 ±0. 170） % ] shown no significant difference comparing to that of control [ （0.20±0.03 ）% ], but the percentage of IFN-γ secreting CD8^＋ T cells in the experimental group （0.28 ± 0.17 % ） was much higher than that of the control[ （0.24 ± 0.12） % ] （ P 〈 0.05, t = 4. 167）. Conclusion The therapeutic HBV DNA vaccine can induce HBsAg specific immune response of IFN-γ secreting cells in which the active CD8^＋ T cells are more predominant.

关 键 词：HBV DNA疫苗 酶联免疫斑点法 流式细胞术 Γ干扰素 

分 类 号：R392[医药卫生—免疫学]