耻垢分枝杆菌与结核分枝杆菌抗原的交叉免疫反应研究

发布时间：2018-01-13 03:34

本文关键词：耻垢分枝杆菌与结核分枝杆菌抗原的交叉免疫反应研究　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的探讨耻垢分枝杆菌(Mycobacteria smegmatis,MS)与结核分枝杆菌(Mycobacteria tuberculosis,MTB)共同抗原及其免疫原性,为耻垢分枝杆菌作为结核潜在疫苗的研究提供科学依据。方法以低温超声破碎、活菌碾磨破碎两种方法提取MS和MTB菌体抗原,以及自己收集的培养后期上清滤液蛋白,通过Western blot观察菌体抗原与抗血清的免疫交叉反应。将MS超声破碎抗原和MTB超声破碎抗原分别与不完全弗氏佐剂混匀后肌肉注射于小鼠,免疫不同次数后7天,以Elispot技术检测小鼠脾脏淋巴细胞经不同细菌抗原分别刺激后能产生抗原特异性IFN-γ的淋巴细胞水平,流式细胞学技术检测各类细胞因子水平,ELISA技术检测血清中抗体效价水平。结果Western blot结果显示MS与MTB间存在一些主要分布在10-100KD的共同抗原,能与两细菌的抗血清产生较强的交叉免疫反应;ELISPOT及流式细胞学结果显示经两细菌分别致敏的小鼠均能两种分枝杆菌抗原产生以细胞免疫为主的免疫应答,随免疫次数的增加免疫反应也增强,免疫两针接近高峰,免疫三针达到平衡并出现机体免疫系统的平衡调节。CD4+T及CD8+T淋巴细胞分泌IL-2、IFN-γ水平均明显高于未免疫组(P0.05),且对非分枝杆菌属的布氏杆菌对照抗原几乎不产生细胞免疫应答。ELISA结果显示两细菌抗原分别免疫的小鼠血清中含有大量能与共同抗原特异性结合的抗体,并且随免疫次数的增加而增高,免疫三针接近高峰。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反应,为MS用于结核新疫苗佐剂等研究提供科学依据。
[Abstract]:Objective to investigate the Mycobacterium smegmatis (Mycobacteria smegmatis, MS) and Mycobacterium tuberculosis (Mycobacteria tuberculosis MTB) common antigen and its immunogenicity as Mycobacterium smegmatis as potential vaccine of tuberculosis and provide scientific basis. Methods with low temperature ultrasonic crushing, milling and extraction of MS bacteria MTB antigen broken two methods. And his collection of late culture filtrate proteins, observe the immune cross reaction antigen and antiserum by Western blot. The MS ultrasonic antigen and MTB supersonic antigen respectively with incomplete Freund's adjuvant mixed intramuscular injection in mice 7 days after immunization, at different times, in order to detect the spleen lymphocytes of mice by different Elispot technology bacterial antigens were stimulated to produce antigen-specific IFN- lymphocyte gamma level, flow cytometry detection of cytokines level of ELISA. Antibody titers were detected in serum. The results showed that some Western blot mainly distributed in the common antigen of the existence of 10-100KD between MTB and MS, can produce strong cross immune response and antibody two bacteria; ELISPOT and flow cytometry analysis showed that the two bacteria were sensitized mice to the immune response to cell immunity is the main two kinds of mycobacterial antigens, the immune response increased with immunization times also enhanced immune, two needle close to the peak, three needle balance and immune immune system regulates the balance of.CD4+T and CD8+T lymphocytes secreting IL-2, IFN- gamma average water was significantly higher than that in non immune group (P0.05), and almost no cellular immune antigen the results showed that serum.ELISA response in mice of two bacterial antigen, contains a large number can be combined with the common antigen of Brucella non Mycobacterium control The antibody increased with the increase of immunization times. The three immunization needles approached the peak. Conclusion these common antigens exist between MS and MTB, so that MS sensitized animals can cause specific immune responses against Mycobacterium tuberculosis, and provide scientific basis for MS in the research of new vaccine adjuvant for tuberculosis.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R392

【参考文献】