免疫蛋白质组学方法对内脏利什曼病人血清识别主要抗原的鉴定

发布时间：2018-07-16 13:04

【摘要】：目的鉴定我国内脏利什曼病人血清识别的主要抗原蛋白。方法培养杜氏利什曼原虫前鞭毛体并体外转化成无鞭毛体,提取总蛋白,经2-DE电泳,以内脏利什曼病人血清为一抗进行2-D Western blot,对强免疫识别点相应抗原蛋白进行MALDI-TOF/TOF串联质谱鉴定。结果等量的杜氏利什曼原虫前鞭毛体与无鞭毛体蛋白经2-DE电泳均可呈现约700个蛋白点,内脏利什曼病人血清识别的主要无鞭毛体抗原蛋白为β-微管蛋白、磷酸多糖β-1,3半乳糖基转移酶、PA7为磷酸多糖β-1,3阿拉伯糖基转移酶及延伸因子1等,识别的主要前鞭毛体抗原蛋白为热休克蛋白70及延伸因子1等。结论人内脏利什曼病患者血清能识别多种杜氏利什曼原虫蛋白,有利于筛选抗内脏利什曼病疫苗抗原和新的血清学诊断抗原。
[Abstract]:Objective to identify the major antigenic proteins recognized by visceral Leishman patients in China. Methods the proflagellum of Leishmania Dursoni was cultured and transformed into flagellum in vitro. The total protein was extracted by 2-DE electrophoresis. The serum of visceral leishmaniacs was used as the first antibody to carry out 2-D Western blot. The corresponding antigen proteins at the strong immunological recognition point were identified by MALDI-TOF / TOF tandem mass spectrometry (MALDI-TOF / TOF tandem mass spectrometry). Results the same amount of proflagellum and free flagellum proteins of Leishmania donovani showed about 700 protein spots by 2-DE electrophoresis. The main antigenic proteins recognized by visceral leishmania patients were 尾 -tubulin, 尾-tubulin, and 尾-tubulin. Phosphopolysaccharide 尾 -1t3 galactosyltransferase (PA7) is phosphopolysaccharide 尾 -1t3 arabinyltransferase and extended-factor 1, and the major proflagellum antigen proteins recognized are heat shock protein 70 and extension factor 1, etc. Conclusion the sera of patients with visceral leishmaniasis can recognize many kinds of Leishmania dunsoni proteins, which is helpful for screening the antigens against visceral leishmaniasis vaccine and new serological diagnostic antigens.
【作者单位】：川北医学院免疫学与分子生物学研究所;
【基金】：国家自然科学基金项目(No.30872213) 四川省应用基础项目(No.2008GY0088-1)
【分类号】：R531.6

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