二甲基三十六烷基铵-卡介苗多糖核酸在结核亚单位疫苗中的佐剂效应  

Dimo-thylidioctyl ammonium bromide-BCG polysaccharide nucleic acid adjuvant enhanced the immunogenicity of a Mycobacterium tuberculosis subunit vaccine

作　　者：宋楠楠[1] 王秉翔[2] 史大中[1] 傅林锋[2] 雒彧[1] 于红娟[1] 韩少波[2] 焦磊[2] 郄亚卿[3] 王洪海[3] 张颖 祝秉东[1] 

机构地区：[1]兰州大学结核病研究中心暨病原生物学研究所,730000 [2]兰州生物制品研究所 [3]复旦大学遗传学研究所 [4]Department ofMolecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University

出　　处：《中华结核和呼吸杂志》2009年第7期513-516,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：基金项目：国家高技术研究发展计划（863计划）资助项目（2006AA02ZA20）;科技重大专项课题资助项目（2008ZX10003-013）

摘　　要：目的探讨二甲基三十六烷基铵（DDA）、卡介苗多糖核酸（BCG—PSN）作为结核病融合蛋白疫苗佐剂的免疫效应。方法采用热酚法制备BCG—PSN，与DDA联合作为融合蛋白AMM（Ag85B—MPT64 190-196-Mtb8．4）佐剂免疫小鼠，设卡介苗、生理盐水、弗氏不完全佐剂（IFA）及单独应用DDA或BCG—PSN一种佐剂的各组作为对照。应用ELISA和酶联免疫斑点法检测免疫小鼠的体液与细胞免疫反应。结果分离培养疫苗免疫小鼠的脾脏淋巴细胞，用特异性抗原Ag85B刺激后，AMM＋DDA＋BCG—PSN疫苗组、AMM＋DDA组和卡介苗组小鼠每1×10^6个脾淋巴细胞分泌γ-干扰素的细胞数分别为222±79、259±85和230±64，均明显高于AMM＋PBS组（40±4）、AMM＋IFA组（10±3）、AMM＋BCG．PSN组（132±18）和生理盐水组（8±4），差异有统计学意义（t值为2．923—7．118，均P〈0．05）；与单用DDA组比较，添加DDA和BCG—PSN两种佐剂的疫苗组小鼠血清抗体滴度Ig2／IgG，较高（0．125和0．025），但γ-干扰素分泌水平没有增高。结论AMM＋DDA＋BCG—PSN结核病亚单位疫苗能够引起较强的Th1细胞免疫为主的免疫反应，DDA＋BCG—PSN（尤其是DDA）具有增强结核病亚单位疫苗Th1细胞免疫应答的佐剂效应。Objective To investigate the activity of a novel adjuvant consisting of dimethyldioctyldecyl ammonium bromide （DDA） and BCG polysaccharide nucleic acid （BCG-PSN）. Methods BCG-PSN was extracted by hot-phenol method, and combined with DDA and Mycobacterium tuberculosis fusion antigen AMM （Ag85B-MPT64 190-198-Mtb8.4） to formulate the Mycobacterium tuberculosis subunit vaccine. Mice were immunized subcutaneously with a 2-week interval between the immunizations （0. 2 ml/ dose）, and humoral and cell-mediated immunity were detected by ELISA and ELISPOT respectively. Results With the stimulation of Ag85B in vitro, the number of antigen specific IFN-γproducing spleen lymphocytes were 222 -＋ 79,259 ± 85,230 ± 64 per million respectively in the mice immunized with AMM ＋ DDA ＋ BCG- PSN, AMM ＋ DDA, and BCG. Spleen lymphocytes in these 3 groups produced higher levels of IFN-γ compared to the groups with the adjuvant of IFA or BCG-PSN alone or without adjuvant upon stimulation with Ag85B （t = 2. 923 -7. 118, P 〈 0. 05）. Furthermore, the adjuvant consisting of DDA and BCG-PSN increased the ratio of Ig2a/IgG1 than DDA alone （ 0. 125 vs 0. 025 ）. Combined with AMM, the adjuvant DDA and the one consisting of DDA and BCG-PSN induced higher level of immunity than incomplete Freund＇s adjuvant （IFA）, NaCl, and BCG-PSN alone. Conclusion Mycobacterium tuberculosis subunit vaccine AMM ＋ DDA ＋ BCG-PSN induced a strong Thl-type immune response, and DDA ＋ BCG-PSN, especially DDA promoted the immune response of the M. tuberculosis subunit vaccine in mice.

关 键 词：分枝杆菌 结核 疫苗 亚单位 卡介苗 

分 类 号：R686[医药卫生—骨科学]