日本血吸虫尾蚴及童虫可溶性抗原蛋白质组研究  

The soluble antigenic proteome of Schistosoma japonicum cercariae and schistosmnula MA An

作　　者：马安[1] 王越[1] 汤益[2] 杨再峰[3] 施晓华[1] 朱明东[1] 刘晓龙[1] 干小仙[1] 

机构地区：[1]浙江省医学科学院寄生虫病研究所,杭州310013 [2]杭州市疾病预防控制中心地方病寄生虫病防治所 [3]浙江省医学科技教育发展中心,杭州310006

出　　处：《国际流行病学传染病学杂志》2011年第4期229-234,共6页International Journal of Epidemiology and Infectious Disease

基　　金：浙江省科技厅专项基金（2007F10029）

摘　　要：目的应用免疫蛋白质组研究方法筛选、鉴定日本血吸虫尾蚴、童虫可溶性抗原蛋白质。方法日本血吸虫尾蚴可溶性粗抗原（scAP）、童虫可溶性粗抗原（SLAP）分别用双向凝胶电泳（2-DE）分离蛋白质，每样本同时制3块胶，1块胶进行银染，2块胶通过电转印后再分别用感染兔和正常兔血清作Western印迹分析，确定特异性阳性反应点，再从相应银染胶图上找到匹配的抗原蛋白质点；用MALDI．TOWTOF串联质谱鉴定抗原蛋白质。结果SCAP、SLAP与感染兔血清免疫反应分别获得94个和68个阳性点，在相应的银染胶图上分别获得33个和31个匹配蛋白质点；用MALDI．TOF／TOF串联质谱鉴定和NCBI数据库检索，鉴定成功率分别为100．O％（20／20）和68．2％（15／22）。已获鉴定的SCAP抗原中蛋白酶占62．5％（10／16），SLAP抗原中蛋白酶占36．4％（4／11）；2个抗原蛋白为SCAP和SLAP共有。结论2-DE能有效地分离日本血吸虫可溶性抗原蛋白质，2-DWestern印迹法能较好地筛选特异性抗原；2-DWestern印迹法阳性点与2-DE胶图蛋白质点匹配率低，低丰度的抗原蛋白质易被漏检；尾蚴和童虫的抗原蛋白质组差异较大。Objective To screen and identify specific antigenic proteins of Schistosomajaponicum cercariae and schistosomula using immunproteomics approaches. Methods Soluble antigenic proteins of Schistosoma japonicum cer- carie （SCAP） and 15 days lung-stage schistosomulum （SLAP） were separated by two-dimensional electropboresis （2- DE）. For each sample, three gels were run in parallel with one gel for silver stain and the other two gels for Western blot using Sch/stosoma japon/cum infected rabbit sera and normal rabbit sera separately. The specific antigenic protein spots were determined on the membrane of Western blot. The matched antigenic protein spots .on the sliver stained gels were subsequently analysed by MALDI-TOF/TOF-MS/MS respectively. Results 94 and 68 positive spots were visualised re- spectively on the membranes of 2-D Western blot of SCAP and SLAP, incubated with Schistosoma japonicum infected rab- bit sera. There were 33 and 31 precisely matched protein spots on the corresponding sliver stained gels of SCAP and SLAP, separately. All SCAP protein spots were identified successfully with MALDI-TOF/TOF-MS/MS and＇ NCBI database retrieval while 68.2% （15/22） of SLAP were confirmed. 62.5% of antigenic proteins of SCAP were protease and that of SLAP was 36.4%. Two antigenic proteins existed both in SCLP and SLAP. Conclusions 2-DE could efficiently separate proteins of SCAP and SLAP and 2-D Western blot could screen specific antigens very well, but the matching rate between positive spots on 2-D Western blot and protein spots on 2-DE silver stained gels were low, and low-abundant antigenic proteins were easily missed to be detected. The antigenic proteomes were significantly different between SCAP and SLAP.

关 键 词：血吸虫 日本 抗原 蛋白质组 

分 类 号：R392[医药卫生—免疫学]