浅黄分枝杆菌和母牛分枝杆菌感染鼠的细胞免疫变化规律研究

发布时间：2018-03-02 22:12

本文选题：浅黄分枝杆菌　切入点：母牛分枝杆菌　出处：《吉林农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的:利用母牛分枝杆菌和浅黄分枝杆菌建立小鼠非结核分枝杆菌持续感染模型。分析感染小鼠体内的免疫细胞、细胞因子和组织病理随感染时间的动态变化。从而为探索非结核分枝杆菌感染的免疫学机制奠定基础。方法:选取雌性C57BL/6小鼠,经腹腔注射感染母牛分枝杆菌和浅黄分枝杆菌。分别于感染后2、7、14、21、28、35天取材,称量肝脏、脾脏、肺脏重量并制作病理切片经HE染色观察感染小鼠器官的病理变化;通过荧光定量PCR检测肝脏和脾脏中IFN-γ、TNF-α、TGF-β、NLRP3、IL-12b和IL-10 6种细胞因子mRNA水平表达量的变化;采用流式细胞术检测脾脏CD3+、CD4+、CD8+T淋巴细胞的细胞比例。结果:1、经母牛分枝杆菌感染的小鼠在整个感染过程中,肝脏和肺脏可见明显的病理变化,感染后第7天,肝脏中这6种细胞因子的mRNA水平表达量达到了最低点,随后逐渐增加一直到第28天,脾脏仅诱导微弱的IL-10、TNF-α、IL-12b和TGF-β的表达。CD4+T淋巴细胞的比例先降低后升高,CD8+T淋巴细胞的比例先升高后降低。2、经浅黄分枝杆菌感染的小鼠在整个感染过程中,肝脏、脾脏和肺脏均可见微弱的病理变化。肝和脾中IFN-γmRNA的表达逐渐增加。IL-12b在感染初期阶段持续降低,感染后第14天降至最低,随后逐渐升高。肝脏中IL-10的表达在感染后21天达到峰值,随后逐渐降低,而脾脏中IL-10在感染后14天达到峰值,之后显著降低。肝脏和脾脏中只诱导微弱的NLRP3、TNF-α和TGF-βmRNA的表达。CD4+T淋巴细胞的比例先降低后升高,CD8+T淋巴细胞的比例先升高后降低。结论:1、母牛分枝杆菌感染组,小鼠肝脏中IFN-γ、IL-12b、IL-10、NLRP3、TNF-α和TGF-β6种细胞因子的表达量均显著高于脾脏,且肝脏显示明显的病理变化,表明肝脏的炎症反应比脾脏更强烈。2、浅黄分枝杆菌感染组,小鼠肝脏和脾脏内IFN-γ、IL-12b和IL-10均发挥明显的免疫效应,而NLRP3、TNF-α和TGF-β的表达量较低。3、CD4+、CD8+T淋巴细胞在非结核分枝杆菌感染过程中均发挥重要的免疫效应。
[Abstract]:Objective: to establish a continuous infection model of non-tuberculous mycobacterium in mice by using Mycobacterium cow and Mycobacterium palliformis, and to analyze the immune cells in infected mice. The dynamic changes of cytokines and histopathology with infection time laid a foundation for exploring the immunological mechanism of non-tuberculous Mycobacterium tuberculosis infection. Methods: female C57BL / 6 mice were selected. The infected mice were infected with Mycobacterium cow and Mycobacterium palliformis by intraperitoneal injection. The samples were collected on the 2nd day after infection, weighing the weight of liver, spleen and lung, and making pathological sections to observe the pathological changes of infected mouse organs by HE staining. The expression of TNF- 伪 TGF- 尾 -NLRP3IL-12b and IL-10 in liver and spleen were detected by fluorescence quantitative PCR. Flow cytometry was used to detect the percentage of CD4 and CD8 T lymphocytes in spleen. Results: the pathological changes of liver and lung were observed in mice infected with Mycobacterium vaccinalis during the whole process of infection, and the pathological changes were observed on the 7th day after infection. The expression of mRNA in the liver reached its lowest level, and then increased gradually until the 28th day. The percentage of IL-10 TNF- 伪 -IL-12b and TGF- 尾. CD4 T lymphocytes decreased first, then increased, and then decreased. 2. The liver of mice infected with Mycobacterium palliformis was observed in the whole process of infection. The expression of IFN- 纬 mRNA in liver and spleen increased gradually in the early stage of infection, decreased to the lowest on the 14th day after infection, and then increased gradually. The expression of IL-10 in liver reached its peak at 21 days after infection. Then decreased gradually, and the IL-10 in spleen reached a peak at 14 days after infection. The percentage of TNF- 伪 and TGF- 尾 mRNA in liver and spleen decreased first, then increased and then decreased. The expression of TNF- 伪 and TGF- 尾 in the liver of mice was significantly higher than that in the spleen, indicating that the inflammatory reaction of the liver was stronger than that of the spleen, and the inflammatory response of the liver was much stronger than that of the spleen, while the expression of TNF- 伪 and TGF- 尾 in the liver was significantly higher than that in the spleen, which indicated that the inflammatory reaction of the liver was stronger than that of the spleen. The expression of IFN- 纬 IL-12b and IL-10 in the liver and spleen of mice showed obvious immune effects, while the expression of NLRP3TNF- 伪 and TGF- 尾 was lower. The expression of CD4 / CD8 T cells in the liver and spleen of mice played an important immune effect in the process of infection with non-tuberculous Mycobacterium tuberculosis.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.4

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