结核病家兔皮肤病理模型与免疫病理机制研究

发布时间：2018-07-04 13:49

本文选题：结核 + 病理　；参考：《兰州大学》2009年硕士论文

【摘要】： 第一部分:分枝杆菌所致家兔皮肤液化病理模型研究研究目的皮内接种免疫建立BCG、H37Ra和耻垢分枝杆菌感染的新西兰兔皮肤模型,为肺结核干酪样坏死和继而发生的液化提供研究模型。材料和方法将三种菌株卡介苗(Bacillus Calmette-Guerin,BCG),耻垢分枝杆菌(M.smegmatis),H37Ra株配置成5×106CFU、5×104CFU、5×102CFU菌液。对健康新西兰兔初次免疫分别行皮内注射5×106CFU、5×104CFU、5×102CFU的BCG、H37Ra和耻垢分枝杆菌菌液。6周后在病灶周围再次以相同剂量行皮内注射进行加强免疫,加强免疫后14天后病变明显时取材,以石蜡包埋,制作切片并行HE染色于镜下观察。结果新西兰兔分别经皮内接种BCG、H37Ra和耻垢分枝杆菌后,高剂量组观察到明显的炎症反应和脓肿液化、破溃等病理改变。再次免疫后可观察到Koch现象。引起病变的严重程度依次为BCG强于H37Ra,后者又强于耻垢分枝杆菌。皮肤模型处取材行细菌抗酸染色,结果呈阳性。三组病灶切片的显微病理改变均可观察到典型的结核结节样病灶。BCG中、低剂量组再次免疫后可诱导产生小结节样改变,但不发生液化溃疡,H37Ra中剂量组和耻垢分枝杆菌中剂量组及所有低剂量组均没有观察到明显的病理改变。结论分枝杆菌引起的皮肤干酪样坏死和液化与感染细菌剂量密切相关,5X106CFU/ml浓度的分枝杆菌可有效诱导新西兰兔皮肤病灶发生液化和坏死,其中BCG引起的病理改变最明显。第二部分:人空洞型肺结核病理组织CD4、CD8、CD25和CD68分子表达分析研究目的应用免疫组织化学技术检测慢性纤维空洞型肺结核病人肺病理组织中CD4~+T、CD8~+T、CD25~+T淋巴细胞和CD68~+巨噬细胞的增殖情况及分布特点,探究肺结核发生发展的免疫机制。材料和方法兰州市肺科医院2005至2006年肺结核病人手术标本中选取的10例石蜡标本。应用免疫组织化学两步法检测10例空洞性肺结核干酪样坏死组织石蜡切片中CD4、CD8、CD25和CD68阳性细胞分布特征。结果在空洞性结核为主的结核组织中,以坏死部位为中心,各种免疫细胞出现了明显的分层分布和在坏死外周出现了大量的细胞浸润。在外周细胞浸润细胞中,CD25+T淋巴细胞为中心CD4~+ T和CDS~+ T细胞聚集,CD68+巨噬细胞也少量聚集。结论外周细胞浸润处以CD25+ T淋巴细胞为中心的细胞浸润区域是宿主病原菌相互作用的免疫活性中心。
[Abstract]:Part one: study on the pathological model of skin liquefaction induced by mycobacterium in rabbits objective to establish the skin model of New Zealand rabbits infected by BCGH37Ra and Mycobacterium smeareus by intradermal immunization. To provide a research model for caseous necrosis and subsequent liquefaction of pulmonary tuberculosis. Materials and methods three strains of Bacillus Calmette-Guerinen BCG and M.smegmatis H37Ra strain were divided into 5 脳 10 6 CFU 5 脳 10 4 CFU 5 脳 10 2 CFU solution. Healthy New Zealand rabbits were immunized by intradermal injection of 5 脳 10 6 CFU 5 脳 10 4 CFU 5 脳 10 2 CFU 5 脳 10 2 CFU respectively. After 6 weeks, the rabbits were immunized again with the same dose of intradermal injection around the lesion. Paraffin embedded sections were made and stained with HE under microscope. Results after intradermal inoculation of BCGH37Ra and Mycobacterium smegmatis in New Zealand rabbits, obvious inflammatory reaction, liquefaction and rupture of abscess were observed in high dose group. Koch phenomenon was observed after reimmunization. The severity of BCG was stronger than that of H37 Ra.The latter was stronger than Mycobacterium smearus. Bacterial acid-fast staining was performed on the skin model and the results were positive. In the typical tuberculous nodular lesion. BCG was observed in the micropathological changes of the lesion sections in the three groups. The small nodular changes could be induced in the low dose group after repeated immunization. However, no significant pathological changes were observed in the medium dose group of H37Ra, the medium dose group of Mycobacterium smearus and all the low dose groups. Conclusion the keratinoid necrosis and liquefaction caused by mycobacterium can induce liquefaction and necrosis of skin lesions in New Zealand rabbits with the concentration of 5X106 CFU / ml. The pathological changes caused by BCG are the most obvious. The second part: expression of CD4, CD8, CD25 and CD68 in human cavitary pulmonary tuberculosis objective to detect the expression of CD4 ~ T ~ + CD8 ~ in lung tissues of patients with chronic fibrous cavitation pulmonary tuberculosis by immunohistochemical technique The proliferation and distribution of CD25 ~ T lymphocytes and CD68 ~ macrophages. To explore the immune mechanism for the occurrence and development of pulmonary tuberculosis. Materials and methods Ten paraffin specimens from pulmonary tuberculosis patients from 2005 to 2006 were selected. Immunohistochemical two-step method was used to detect the distribution of CD4 + CD8 + CD25 and CD68 positive cells in paraffin sections of 10 caseous necrotic tissues of pulmonary tuberculosis. Results in the tuberculous tissues with cavitary tuberculosis, the necrotic site was the center, and the various immune cells were obviously stratified and infiltrated in the necrotic periphery. In peripheral infiltrating cells, CD25 T lymphocytes were the center of CD4T and CDS- T cells, and CD68 macrophages were also concentrated in a small amount. Conclusion the infiltrating area of peripheral cells with CD25 T lymphocytes as the center is the immune active center of host pathogen interaction.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R52;R392

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