嗜肺军团菌抗原组分及γ干扰素抗其感染的研究
发布时间：2019-06-29 22:58
【摘要】：目的 嗜肺军团菌的抗原组分与常见细菌性肺炎病原菌之间共同抗原的研究:观察IFN-γ增强豚鼠抗嗜肺军团菌感染的作用。 方法 制备1型嗜肺军团菌(LP1)菌体抗原,并用之多次免疫家兔,取其血清(多克隆抗体)作为抗体;提取LP1、肺炎链球菌、流感嗜血杆菌、铜绿假单胞菌、肺炎克雷伯杆菌、金黄色葡萄球菌、表皮葡萄球菌和大肠杆菌的膜抗原;将抗原进行聚丙烯酰胺凝胶电泳(SDS-PAGE);再把凝胶上的蛋白质转移到PVDF膜上;进行免疫印迹分析(Western-blot)。 将LP1经气管感染豚鼠,随机分成两组,治疗组和对照组,前者给予肌肉注射IFN-γ,后者肌肉注射PBS;每天观察两组豚鼠的一般体征;并于感染后的第24h、48、72h、120h和7d各处死4只,分别从腹主静脉抽血分离血清,检测CPK、GOT、GPT、AKP、BUN、Cr和LDH水平;取肺组织进行肺部菌落计数,并观察其肺病理改变。 结果 SDS-PAGE后的凝胶经考马斯亮染色显示,LP1膜抗原的主要条带位于154kDa、97kDa、79kDa、58kDa和44kDa处,其结果在Western-blot分析上得到了证实。Western-blot结果显示:约在154kDa和97kDa处表皮葡萄球菌、流感嗜血杆菌和铜绿假单胞菌泳道上同样可见清晰的条带;在79kDa处流感嗜血杆菌、铜绿假单胞菌泳道上也有清晰的条带;在58kDa处大肠杆菌、流感嗜血杆菌、肺炎克雷伯杆菌泳道上也可见有相应的条带;在44kDa处铜绿假单胞菌泳道上也有清晰的条带。金黄色葡萄球菌和肺炎链球菌膜抗原未见任何条带出现。 IFN-γ治疗组动物感染的体征较轻,肺菌落数明显减少,部分感染动物的血清CPK、GOT、AKP、BUN、Cr和LDH水平明显降低,病理改变较对照组明显减轻。 结论 流感嗜血杆菌、铜绿假单胞菌和LP1间存在有三个共同的膜抗原表位;表皮葡萄球菌有两个,大肠杆菌、肺炎克雷伯杆菌只有一个。金黄色葡萄球和肺炎链球菌与
[Abstract]:Objective to study the common antigen between the antigen components of Legionella pneumophilia and the common pathogenic bacteria of bacterial pneumonia: to observe the effect of IFN- 纬 on the anti-Legionella pneumophilia infection in guinea pigs. Methods Legionella pneumophilia type 1 (LP1) antigen was prepared and used to immunize rabbits for many times, and its serum (polyclonal antibody) was taken as antibody, the membrane antigens of Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli were extracted, and polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Then the protein on the gel was transferred to PVDF membrane, and immunoblotting analysis (Western-blot) was performed. The guinea pigs infected with LP1 through trachea were randomly divided into two groups: treatment group and control group, the former group was given intramuscular injection of IFN- 纬, the latter was intramuscular injection of PBS; to observe the general signs of the two groups every day, and 4 guinea pigs died at 24 h, 48 h, 72 h, 120 h and 7 d after infection. Serum samples were taken from abdominal main vein to detect the levels of CPK,GOT,GPT,AKP,BUN,Cr and LDH. The lung colonies were counted and the pathological changes of the lungs were observed. Results the gel after SDS-PAGE was stained by Coomassie bright staining, the main bands of LP1 membrane antigen were 154kDa, 97kDa, 79kDa. the results were confirmed by Western-blot analysis. Western-blot results showed that clear bands were also found in Staphylococcus epidermidis, Haemophilus influenzae and Pseudomonas aeruginosa in 154kDa and 97kDa, and there were also clear bands in Haemophilus influenzae and Pseudomonas aeruginosa in 79kDa. There were also corresponding bands in E. coli, Haemophilus influenzae and Klebsiella pneumoniae in 58kDa, and clear bands in Pseudomonas aeruginosa lane in 44kDa. No bands were found in Staphylococcus aureus and Streptococcus pneumoniae membrane antigen. In the IFN- 纬 treatment group, the signs of infection were mild, the number of lung colonies was significantly decreased, the levels of serum CPK,GOT,AKP,BUN,Cr and LDH in some infected animals were significantly decreased, and the pathological changes were significantly reduced compared with the control group. Conclusion there are three common membrane antigen epitopes between Haemophilus influenzae, Pseudomonas aeruginosa and LP1, and there are only one in Staphylococcus epidermidis and only one in Escherichia coli and Klebsiella pneumoniae. Staphylococcus aureus and Streptococcus pneumoniae
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R378
本文编号：2508191
[Abstract]:Objective to study the common antigen between the antigen components of Legionella pneumophilia and the common pathogenic bacteria of bacterial pneumonia: to observe the effect of IFN- 纬 on the anti-Legionella pneumophilia infection in guinea pigs. Methods Legionella pneumophilia type 1 (LP1) antigen was prepared and used to immunize rabbits for many times, and its serum (polyclonal antibody) was taken as antibody, the membrane antigens of Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli were extracted, and polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Then the protein on the gel was transferred to PVDF membrane, and immunoblotting analysis (Western-blot) was performed. The guinea pigs infected with LP1 through trachea were randomly divided into two groups: treatment group and control group, the former group was given intramuscular injection of IFN- 纬, the latter was intramuscular injection of PBS; to observe the general signs of the two groups every day, and 4 guinea pigs died at 24 h, 48 h, 72 h, 120 h and 7 d after infection. Serum samples were taken from abdominal main vein to detect the levels of CPK,GOT,GPT,AKP,BUN,Cr and LDH. The lung colonies were counted and the pathological changes of the lungs were observed. Results the gel after SDS-PAGE was stained by Coomassie bright staining, the main bands of LP1 membrane antigen were 154kDa, 97kDa, 79kDa. the results were confirmed by Western-blot analysis. Western-blot results showed that clear bands were also found in Staphylococcus epidermidis, Haemophilus influenzae and Pseudomonas aeruginosa in 154kDa and 97kDa, and there were also clear bands in Haemophilus influenzae and Pseudomonas aeruginosa in 79kDa. There were also corresponding bands in E. coli, Haemophilus influenzae and Klebsiella pneumoniae in 58kDa, and clear bands in Pseudomonas aeruginosa lane in 44kDa. No bands were found in Staphylococcus aureus and Streptococcus pneumoniae membrane antigen. In the IFN- 纬 treatment group, the signs of infection were mild, the number of lung colonies was significantly decreased, the levels of serum CPK,GOT,AKP,BUN,Cr and LDH in some infected animals were significantly decreased, and the pathological changes were significantly reduced compared with the control group. Conclusion there are three common membrane antigen epitopes between Haemophilus influenzae, Pseudomonas aeruginosa and LP1, and there are only one in Staphylococcus epidermidis and only one in Escherichia coli and Klebsiella pneumoniae. Staphylococcus aureus and Streptococcus pneumoniae
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R378
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