Comparative analysis of excretory-secretory antigens of Trichinella spiralis and Trichinella britovi muscle larvae by two-dimensional difference gel electrophoresis and immunoblotting.
Comparative analysis of excretory-secretory antigens of Trichinella spiralis and Trichinella britovi muscle larvae by two-dimensional difference gel electrophoresis and immunoblotting.
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DOI：
10.1186/1477-5956-10-10
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发表时间：
2012-02-11
期刊：
影响因子：
2
通讯作者：
Savijoki K
中科院分区：
文献类型：
--
作者：
Bien J;Näreaho A;Varmanen P;Gozdzik K;Moskwa B;Cabaj W;Nyman TA;Savijoki K
旋毛虫病是由旋毛虫属引起的人畜共患疾病。本研究旨在从T. spiralis和T. britovi肌幼虫（ML），有望用于物种特异性诊断。为此，纯化的E-S蛋白进行了分析，通过荧光二维差异凝胶电泳（2-D DIGE）结合蛋白质鉴定液相色谱-串联质谱（LC-MS/MS）。为了寻找能被宿主抗体特异性识别的免疫反应性蛋白质，将E-S蛋白质与来自实验感染T. spiralis或T.布里托维。根据2-D DIGE分析，总共22种蛋白质，包括潜在的免疫原性蛋白质和仅由两种旋毛虫中的一种产生的蛋白质，进行LC-MS/MS用于蛋白质鉴定。从这些蛋白质中可以鉴定出17种，其中许多在多个点中鉴定，表明它们经历了翻译后修饰，可能涉及糖基化和/或蛋白水解。这些蛋白质包括5 '-核苷酸酶、丝氨酸型蛋白酶/蛋白酶、p43糖蛋白（gp 43）以及49 kDa E-S蛋白（p49）。我们的研究结果还表明，一些常见的蛋白质在不同程度上进行了后修饰，在某些情况下，这似乎会导致物种特异性修饰。通过2-DE免疫印迹鉴定了共同和特异性识别的免疫反应蛋白;共同抗原被鉴定为gp 43和不同的蛋白酶变体，而特异于T. britovi包括5 ′-核苷酸酶的多种同种型。二维DIGE和二维免疫印迹法均表明T. spiralis和T. britovi产生一些独特的抗原谱，其中包含E-S抗原，具有作为旋毛虫属种特异性诊断标志物的潜力。我们的研究结果也表明了价值的2-D DIGE作为一个多功能的工具，比较不同的旋毛虫物种的分泌物，查明因素与主机的相互作用。
Trichinellosis is a zoonotic disease in humans caused by Trichinella spp. The present study was undertaken to discover excretory-secretory (E-S) proteins from T. spiralis and T. britovi muscle larvae (ML) that hold promise for species-specific diagnostics. To that end, the purified E-S proteins were analyzed by fluorescent two-dimensional difference gel electrophoresis (2-D DIGE) coupled with protein identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS). To search for immunoreactive proteins that are specifically recognized by host antibodies the E-S proteins were subjected to two-dimensional (2-DE) immunoblotting with antisera derived from pigs experimentally infected with T. spiralis or T. britovi. According to 2-D DIGE analysis, a total of twenty-two proteins including potentially immunogenic proteins and proteins produced only by one of the two Trichinella species were subjected to LC-MS/MS for protein identification. From these proteins seventeen could be identified, of which many were identified in multiple spots, suggesting that they have undergone post-translational modification, possibly involving glycosylation and/or proteolysis. These proteins included 5'-nucleotidase, serine-type protease/proteinase, and p43 glycoprotein (gp43) as well as 49 kDa E-S protein (p49). Our findings also suggest that some of the commonly identified proteins were post-translationally modified to different extents, which in certain cases seemed to result in species-specific modification. Both commonly and specifically recognized immunoreactive proteins were identified by 2-DE immunoblotting; shared antigens were identified as gp43 and different protease variants, whereas those specific to T. britovi included multiple isoforms of the 5'-nucleotidase. Both 2-D DIGE and 2-DE immunoblotting approaches indicate that T. spiralis and T. britovi produce somewhat distinctive antigen profiles, which contain E-S antigens with potential as species-specific diagnostic markers for Trichinella. Our results also demonstrate the value of 2-D DIGE as a versatile tool to compare secretomes of different Trichinella species for pinpointing factors contributing to the interaction with the host.
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影响因子：
1.5
作者：
Gounaris, K;Selkirk, ME;Sadeghi, SJ
通讯作者：
Sadeghi, SJ
影响因子：
2.6
作者：
Dea-Ayuela, MA;Bolás-Fernández, F
通讯作者：
Bolás-Fernández, F
DOI：
10.1007/s00018-004-4445-9
发表时间：
2005-03
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Dubin G
通讯作者：
Dubin G
影响因子：
2.4
作者：
Boireau, P;Vayssier, M;Soulé, C
通讯作者：
Soulé, C
影响因子：
2.6
作者：
Boireau, P;Vallée, I;Gajadhar, A
通讯作者：
Gajadhar, A