结核融合蛋白AMM亚单位疫苗强化BCG初始免疫的免疫效应  

The effects of immunity induced by BCG priming and tuberculosis fusion protein AMM vaccine boosting

作　　者：姜雯雯[1,2] 景涛[1,2,6] 于红娟[2,5] 李青[2,4] 易娟 雒彧[2,4] 宋楠楠[1] 张颖[2,7] 祝秉东[1,2,6] 

机构地区：[1]兰州大学病原生物学研究所,730000 [2]兰州大学结核病研究中心,730000 [3]兰州大学基础医学院实验中心,730000 [4]兰州大学病理生理学研究所,730000 [5]兰州大学免疫学研究所,730000 [6]甘肃省新药临床前研究重点实验室,兰州730000 [7]美国霍普金斯大学布隆博格公共卫生学院分子微生物学与免疫学系

出　　处：《中华微生物学和免疫学杂志》2009年第7期631-635,共5页Chinese Journal of Microbiology and Immunology

基　　金：重大传染病防治科技重大专项（2008zx1000301104）;国家科技部863项目（2006AA02Z420）

摘　　要：目的研究结核融合蛋白Ag85B—Mpt64190-198-Mtb8．4（AMM）和佐剂二甲基三十六烷基铵（DDA）、卡介苗多糖核酸（BCG—PSN）构建的亚单位疫苗强化BCG初始免疫的免疫效应。方法将融合蛋白AMM、佐剂DDA和BCG—PSN混合构建AMM亚单位疫苗。实验1组BCG初免后第10周用AMM亚单位疫苗加强免疫小鼠一次；实验2组BCG初免后分别于第8周、第10周用AMM亚单位疫苗加强免疫小鼠一次。同时设立生理盐水及仅BCG免疫两个对照组。BCG初免后第14周、第22周，应用ELISPOT、ELISA检测免疫小鼠的细胞及体液免疫反应。同时在第22周用BCG活菌攻击被免疫小鼠，间隔4周后用流式细胞术和ELISA技术检测T细胞分型及体液免疫反应。结果（1）IFN-γ水平：BCG初免后14周，特异性抗原Ag85B刺激后实验2组分泌IFN-γ的细胞数（135±14）明显高于仅BCG免疫组（19±16），t=10．98，P〈0．01；BCG初免后22周，实验2组（208±11）同样高于仅BCG免疫组（57±18），t=6．43，P〈0．01。（2）体液免疫应答水平：实验2组的IgG1抗体滴度明显高于实验1组，而作为反映Th1型免疫反应指标的IgG2a／IgG1比值，强化免疫两次组低于强化一次组。（3）BCG模拟攻击被免疫小鼠后，CD4^＋CD25^＋调节性T细胞含量：实验1、2组均高于仅BCG免疫组（t1=3．08，t2=3．16，P〈0．05）。结论BCG免疫-AMM亚单位疫苗加强免疫两次能够引起较强的细胞及体液免疫反应，同时激活调节性免疫反应。Objective To investigate the boosting efficiency of a subunit vaccine consisting of the fusion protein Ag85B-Mpt64190-198-Mtb8.4 （AMM） , dimethyl-dioctyldecyl ammonium bromide （DDA） and BCG polysaccharide nucleic acid （BCG-PSN） on the primed inoculation with BCG. Methods The AMM subunit vaccine was composed of fusion protein AMM, adjuvant DDA and BCG-PSN. The first mouse experimental group was immunized with BCG first, then boosted with the AMM subunit vaccine in the 10th week. The second experimental group was boosted with the AMM subunit vaccine in the 8th week and the lOth week respectively with a two weeks interval after the primed with BCG. Two control groups were treated respectively with physiological saline alone and BCG alone. After the primed inoculation, ELISPOT and ELISA were used for the detection of the cell-mediated and humoral immune response in week 14 and week 22 re- spectively. Furthermore, the immunized mice were challenged with live BCG to mimic tuberculosis infection in the 22nd week after the primed inoculation. Subsequently the T cell typing and humoral response were detected by flow cytometry and ELISA, respectively. Results （ 1 ） The level of secreting IFN-γ ： 14 weeks after the primed inoculation ,with the stimulation of the specific antigen-Ag85B, the number of cells secreting IFN-γin the second experimental group （ 135 ± 14） was more than BCG alone immunized group （ 19 ±16）, t = 10.98, P 〈0.01. In the 22nd week, the number of cells secreting IFN-γ in the second experimental group （208 ± 11 ） was still more than BCG alone group （57±18）, t =6.43, P 〈0.01. （2） The level of humoral immune response： the IgG1 antibody titer in the second experimental group was obviously higher than that in the first experimental group. However, the ratio of IgG2a to IgG1, as the index reflecting the Thl-type immune response, in the experimental group 2 was lower than that in the experimental group 1. （ 3 ） The contents of CD4^＋ CD25 ^4 T cells after c

关 键 词：亚单位疫苗 融合蛋白 BCG 免疫策略 加强免疫 

分 类 号：R392[医药卫生—免疫学]