耻垢分枝杆菌与结核分枝杆菌抗原的交叉免疫应答研究  

Cross immune reaction between Mycobacteria smegmatis and Mycobacteria tuberculosis

作　　者：曹俊[1] 卢锦标[2] 谢安平[2] 徐苗[2] 王国治[2] 沈小兵[2] 陈保文[2] 郭述良[1] 

机构地区：[1]重庆医科大学附属第一医院呼吸与危重症学科,400016 [2]中国食品药品检定研究院结核病疫苗室,北京100050

出　　处：《中华微生物学和免疫学杂志》2017年第4期275-280,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目（81570010）

摘　　要：目的探讨耻垢分枝杆菌（Mycobacterla smegmatis，MS）与结核分枝杆菌（Mycobacteria tuberculosis,MTB）抗原及其免疫原性的交叉情况。方法以低温超声破碎方法提取MS和MTB菌体抗原．通过Westernblot观察菌体抗原与抗血清的交叉免疫应答。将超声破碎的菌体抗原与不完全弗氏佐剂混匀后肌肉注射于小鼠，免疫后7d以ELISPOT技术及流式细胞学技术检测小鼠脾脏淋巴细胞经不同细菌抗原分别刺激后产生的各类细胞因子水平，ELISA技术检测血清中抗体效价水平。结果MS与MTB间存在一些交叉抗原。经两细菌致敏的小鼠均能对不同分枝杆菌抗原产生以细胞免疫为主的免疫应答，CIM^＋及CD8^＋T淋巴细胞分泌IL-2、IFN-γ水平均较未免疫组明显增高（P〈0．05），对非分枝杆菌属布氏杆菌的对照抗原几乎不产生细胞免疫应答。结论MS与MTB间存在这些共同抗原使MS致敏动物能引起针对结核分枝杆菌特异性免疫应答，为MS用于结核新疫苗佐剂等研究提供科学依据。Objective To identify the cross-reactive antigens shared by Mycobacteria smegmatis （MS） and Mycobacteria tuberculosis（MTB） and to analyze their antigenicity. Methods Bacterial antigens were extracted from strains of MS and MTB by uhrasonication. Western blot assay was performed to analyze common antigens that reacted with both of the antiserum samples against MS and MTB. The extracted bacte- rial antigens were mixed with incomplete Freund＇s adjuvant and then were injected into muscles of mice. Cytokines secreted by murine spleen lymphocytes following stimulation with various antigens of MS and MTB were determined by ELISPOT and flow cytometry on the 7th day. IgG levels in serum samples were detected by ELISA 7 days after injection. Results There were cross-reactive antigens shared by MS and MTB. Po- tent humoral immune responses and cellular immunity against both MS and MTB could be induced by those cross-reactive antigens after sensitization the mice by either MS or MTB antigens. Cytokines of IL-2 and IFN-γ in CD4^＋ and CD8^＋T cells of mice stimulated with MS or MTB antigens were significantly increased as compared with those of non-sensitization group and those of Brucella antigens stimulation group. Conclusion Cross-reactive antigens shared by MS and MTS can effectively promote specific immune reactions to the infec- tion of MTB, which provides a scientific basis for the develoDment of tuberculosis vaccines.

关 键 词：结核分枝杆菌 耻垢分枝杆菌 交叉抗原 免疫应答 

分 类 号：R392[医药卫生—免疫学]