卡介苗免疫小鼠T淋巴细胞亚群及其内穿孔素的表达及意义

发布时间：2018-11-08 14:05

【摘要】： 目的:探讨卡介苗(BCG)免疫小鼠T淋巴细胞亚群CD3+、CD4+、CD8+及CD4+CD8+(双阳,DP)T细胞及其内穿孔素(PFN)的表达及意义。方法:1.实验动物分组:按体重相近的原则将30只清洁级KM雌性小鼠随机分成2组,每组15只:①卡介苗免疫组(BCG组);②卡介苗免疫成功后致病量结核杆菌攻击组(BCG-TB组).2.取血:实验开始时第①、②组每只小鼠给予BCG 0.3ml(相当于湿菌0.1 mg/只)皮下注射;第3个月后第②组每只小鼠均给予1ml TB混悬液(细菌数109)经尾静脉内注射,同时,对照组①组注入相同体积的生理盐水作对照。饲养1个月后两组每只小鼠均经内眦静脉取血1.5-2ml,分别检测外周血中T细胞亚群及其内穿孔素的表达。3.检测:以CD3PerCP、CD4FTTC、CD8APC单抗标记T细胞亚群,以PE标记PFN,以流式细胞仪检测分析总的淋巴细胞、CD3+、CD4+、CD8+、CD4+ CD8+(DP)T淋巴细胞计数和各亚群占淋巴细胞百分率及观察各T细胞亚群内表达PFN阳性细胞计数和各自占淋巴细胞百分率。4.肺组织病理学和细菌学检查。结果: 1.小鼠获得卡介苗免疫成功后,受到致病量结核菌攻击时不发病. 2.BCG-TB组小鼠CD3+、CD8+、CD8+%、DP T细胞计数高于BCG组(P0.05),CD4+ / CD8+低于BCG组(P0.05),差异有统计学意义。 3.两组小鼠T细胞亚群均能表达PFN. 4.BCG-TB组小鼠表达PFN的CD3+、CD8+、DP T细胞计数高于BCG组(P0.05),CD4+ / CD8+低于BCG组(P0.05),差异有统计学意义。 5.BCG-TB组小鼠肝脾肺组织切片未见结核病灶,抗酸染色未见结核杆菌。结论: (1)BCG免疫可提高宿主抵抗结核病的免疫力,增强抗结核保护性免疫表现为CD8+T、DP T细胞升高及T细胞内PFN表达的升高.(2)CD8+T、DP T细胞是抗结核保护性免疫相关细胞.(3)PFN表达升高是机体抗结核保护性免疫表现之一。
[Abstract]:Aim: to investigate the expression and significance of CD3, CD4, CD8 and CD4 CD8 (double positive, DP) T cells) and perforin (PFN) (PFN) in T lymphocyte subsets of mice immunized with BCG (BCG). Methods: 1. According to the principle of similar body weight, 30 clean KM female mice were randomly divided into 2 groups, 15 in each group: 1 BCG immunization group (BCG group); 2 the pathogenic amount of Mycobacterium tuberculosis after the successful immunization of BCG vaccine (BCG-TB group). 2. Blood collection: at the beginning of the experiment, each mouse in group 1 was injected with BCG 0.3ml (equivalent to 0.1 mg/ of wet bacteria) subcutaneously. After 3 months, each mouse in group 2 was injected with 1ml TB suspension (bacterial number 109) via caudal vein, and the control group 1 was injected with the same volume of normal saline as control group. After feeding for 1 month, the blood samples of each group were collected from the inner canthus vein to detect the expression of T cell subsets and perforin in peripheral blood. Detection: T-cell subsets were labeled with CD3PerCP,CD4FTTC,CD8APC monoclonal antibody and PFN, with PE. Total lymphocytes, CD3, CD4, CD8 were detected by flow cytometry. CD4 CD8 (DP) T lymphocyte count and the percentage of each subgroup in lymphocytes were observed. The expression of PFN positive cells in each T cell subgroup and the percentage of PFN positive cells in each T cell subgroup were observed. 4. Lung histopathology and bacteriological examination. Results: 1. After BCG was successfully immunized, the mice did not develop the disease when attacked by the pathogenic amount of tuberculous bacilli. The cell counts of CD3, CD8, CD8%, DP T in 2.BCG-TB group were higher than those in BCG group (P0.05), CD4 / CD8 was lower than BCG group (P0.05), the difference was statistically significant. 3. T-cell subsets in both groups expressed PFN. The number of CD3 and CD8, DP T cells expressing PFN in 4.BCG-TB group was higher than that in BCG group (P0.05), CD4 / CD8 was lower than BCG group (P0.05), the difference was statistically significant. In 5.BCG-TB group, no tuberculosis foci were found in liver, spleen and lung sections, and no mycobacterium tuberculosis was found in acid-fast staining. Conclusion: (1) BCG immunization can enhance the immunity of host against tuberculosis, and enhance the protective immunity of anti-tuberculosis. The expression of CD8 TDP T cells and the expression of PFN in T cells are increased. (2) the expression of PFN in T cells is increased. DP T cells are associated with antituberculous protective immunity. (3) the increase of PFN expression is one of the features of anti-tuberculosis protective immunity.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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