结核杆菌Ag85B-DNA与H37Ra序贯免疫小鼠后细胞免疫的探讨  

CELLULAR IMMUNITY INDUCED IN MICE BY A MYCOBACTERIUM TUBERCULOSIS DNA-H37RA PRIME-BOOST VACCINATION REGIMEN

作　　者：王锦彤[1] 冉柏林[1] 卢贤瑜[1] 

机构地区：[1]重庆医科大学临床免疫学教研究室,临床检验诊断学省部共建教育部重点实验室,重庆市临床检验诊断学重点实验室,重庆400016

出　　处：《现代预防医学》2007年第11期2015-2017,共3页Modern Preventive Medicine

基　　金：重庆市教委资助项目(渝教科[2004]12号文);重医大学科技创新资助项目(CX200204)

摘　　要：[目的]探讨用结核分枝杆菌Ag85B-DNA(pTB30m)和结核菌H37Ra序贯免疫小鼠后产生的特异性细胞免疫。[方法]用碱裂解法制备Ag85B成熟蛋白的真核表达质粒(pTB30m),初次免疫小鼠2周后,用H37Ra皮内加强免疫(DNA-85B/H37Ra组),同时设定Ag85B-DNA/BCG组、H37Ra组、BCG组及未免疫组。加强免疫4w和8w后,检测小鼠脾淋巴细胞被PPD刺激指数(SI)及其IFN-γ、IL-2的表达水平。[结果](1)酶切鉴定pTB30m所含外源基因片段大小正确,纯度较高。(2)DNA-85B/H37Ra组小鼠脾淋巴细胞的SI均显著高于H37Ra组、BCG组和DNA-85B/BCG组(P﹤0.05);(3)所有免疫组IFN-γ及IL-2表达与未免疫组比较,差异有统计学意义(P﹤0.05),DNA-85B/H37Ra组与DNA-85B/BCG组均高于H37Ra组和BCG组(P﹤0.05),DNA-85B/H37Ra略高于DNA-85B/H37Ra,但差异无统计学意义(P﹥0.05)。[结论]DNA-85B/H37Ra序贯免疫效果略优于DNA-85B/BCG组,明显优于H37Ra组和BCG组。[ Objective] To investigate the specific cellular immunity after sequential immunization with mycobacterium tuberculosis Ag85B-DNA （pTB30m） and mycobacterium tuberculosis H37Ra in mice. [Methods] Plasmid DNA （pTB30m） encoding mature form of Ag85B was prepared. The mice were primed with DNA vaccine and boosted with H37Ra by intragastric administration at 2-week intervals （DNA-85B/H37Ra group） . Meanwhile, the others were divided into DNA-85B /BCG group、H37Ra group、BCG group and naive control group respectively. At various intervals （4 weeks, 8weeks） after sequentim immunization, the spleen lymphocytes of mice were stimulated with PPD in vitro. Lymphocytes transformation test was deteeted by MTT assay, the expression levels of IFN-γand IL-2 in the supernatants of culture were tested by ELISA. [Rosults] （1） Prepared recombinant eukaryotic expressing right vector was confirmed by restriction endonuclease digestion, and it was examined so highly pure; （2） The stimulation index （SI） of spleen lymphocytes in DNA-85B/H37Ra group was statistically higher than that in H37Ra group, BCG group and DNA-85B/BCG group （P 〈 0.05） ; （3） Compared with naive control group, the expression levels of IFN-γ and IL-2 in immunization group were significant different （P 〈 0.05） . The expression levels in DNA-85B /H37Ra group, DNA-85B /BCG group were all higher than those in H37Ra group, BCG group （P 〈 0.05）, and those in DNA-85B/H37Ra group were slightly better than those in DNA-85B/BCG group, but there was no significant difference （P〉 0.05） . [Conclusion] Cellular immunity induced in mice by a DNA-H37Ra prime-boost vaccination regimen is slightly better than DNA-85B/BCG group, and significantly better than H37Ra group and BCG group.

关 键 词：结核分枝杆菌 序贯名疫策略 抗原Ag85B H37RA BCG 

分 类 号：R378.911[医药卫生—病原生物学]