【摘要】：目的 嗜肺軍團菌的抗原組分與常見細菌性肺炎病原菌之間共同抗原的研究:觀察IFN-γ增強豚鼠抗嗜肺軍團菌感染的作用。 方法 制備1型嗜肺軍團菌(LP1)菌體抗原,并用之多次免疫家兔,取其血清(多克隆抗體)作為抗體;提取LP1、肺炎鏈球菌、流感嗜血桿菌、銅綠假單胞菌、肺炎克雷伯桿菌、金黃色葡萄球菌、表皮葡萄球菌和大腸桿菌的膜抗原;將抗原進行聚丙烯酰胺凝膠電泳(SDS-PAGE);再把凝膠上的蛋白質(zhì)轉(zhuǎn)移到PVDF膜上;進行免疫印跡分析(Western-blot)。 將LP1經(jīng)氣管感染豚鼠,隨機分成兩組,治療組和對照組,前者給予肌肉注射IFN-γ,后者肌肉注射PBS;每天觀察兩組豚鼠的一般體征;并于感染后的第24h、48、72h、120h和7d各處死4只,分別從腹主靜脈抽血分離血清,檢測CPK、GOT、GPT、AKP、BUN、Cr和LDH水平;取肺組織進行肺部菌落計數(shù),并觀察其肺病理改變。 結(jié)果 SDS-PAGE后的凝膠經(jīng)考馬斯亮染色顯示,LP1膜抗原的主要條帶位于154kDa、97kDa、79kDa、58kDa和44kDa處,其結(jié)果在Western-blot分析上得到了證實。Western-blot結(jié)果顯示:約在154kDa和97kDa處表皮葡萄球菌、流感嗜血桿菌和銅綠假單胞菌泳道上同樣可見清晰的條帶;在79kDa處流感嗜血桿菌、銅綠假單胞菌泳道上也有清晰的條帶;在58kDa處大腸桿菌、流感嗜血桿菌、肺炎克雷伯桿菌泳道上也可見有相應(yīng)的條帶;在44kDa處銅綠假單胞菌泳道上也有清晰的條帶。金黃色葡萄球菌和肺炎鏈球菌膜抗原未見任何條帶出現(xiàn)。 IFN-γ治療組動物感染的體征較輕,肺菌落數(shù)明顯減少,部分感染動物的血清CPK、GOT、AKP、BUN、Cr和LDH水平明顯降低,病理改變較對照組明顯減輕。 結(jié)論 流感嗜血桿菌、銅綠假單胞菌和LP1間存在有三個共同的膜抗原表位;表皮葡萄球菌有兩個,大腸桿菌、肺炎克雷伯桿菌只有一個。金黃色葡萄球和肺炎鏈球菌與
[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- 緯 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- 緯, 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- 緯 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
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R378

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