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首页> 外文期刊>PLoS One >Immunoproteomic analysis of Trichinella spiralis and Trichinella britovi excretory-secretory muscle larvae proteins recognized by sera from humans infected with Trichinella
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Immunoproteomic analysis of Trichinella spiralis and Trichinella britovi excretory-secretory muscle larvae proteins recognized by sera from humans infected with Trichinella

机译：Trichinella spirlalis和Trichinella Britovi排泄 - 分泌肌肉幼虫蛋白质的免疫蛋白质分析来自人类感染的人类感染的血清

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The present study compares the immunogenic patterns of muscle larvae excretory-secretory proteins (ML E-S) from T . spiralis and T . britovi recognized by Trichinella -infected human sera. Samples were analyzed using two-dimensional electrophoresis (2-DE) coupled with 2D-immunoblot and liquid chromatography-tandem mass spectrometry LC-MS/MS analysis, two ELISA procedures and a confirmatory 1D-immunoblot test. Sera were obtained from nine patients with a history of ingestion of raw or undercooked meat who presented typical clinical manifestations of trichinellosis and from eleven healthy people. Specific anti- Trichinella IgG antibodies were detected in all samples tested with the Home-ELISA kits, but in only four samples for the commercially-available kit. The 1D-immunoblot results indicated that all nine serum samples were positive for T . spiralis ML E-S antigens, expressed as the presence of specific bands. In contrast, eight of the serum samples with T . britovi E-S ML antigens were positive, with one serum sample taken from a patient at 33dpi (days post infection) being negative. To identify immunoreactive proteins that are specifically recognized by host antibodies, both species of ML E-S proteins were subjected to 2D-immunoblotting with human serum taken at 49 dpi. The sera recognized 22 protein spots for T . spiralis and 18 for T . britovi in 2D-immunoblot analysis. Their molecular weights (MW) ranged from 50 to 60 kDa. LC-MS/MS analysis identified both common and specifically-recognized immunoreactive proteins: transmembrane serine protease 9, serine protease, antigen targeted by protective antibodies and Actin-1 partial were shared for both Trichinella species; hypothetical protein T01_7775 and P49 antigen, partial those specific to T . spiralis ; deoxyribonuclease-2-alpha and hypothetical protein T03_17187/T12_7360 were specific to T . britovi . Our results demonstrate the value of 2-DE and 2D-immunblot as versatile tools for pinpointing factors contributing to the parasite-host relationship by comparing the secretomes of different Trichinella species.
机译：本研究比较了肌肉幼虫排泄 - 分泌蛋白（ML E-S）的免疫原性图案。 spiralis和t。 Britovi由Trichinella-infaced人类血清认可。使用二维电泳（2-DE）与2D-IMMUNBLOT和液相色谱 - 串联质谱LC-MS / MS分析，两个ELISA程序和确认1D-IMMUNBLOT试验进行分析样品。血清是从九个患者获得的患有生物或未煮熟的肉类的历史，他们呈现典型的孕粒症和11个健康人群。在用家用ELISA试剂盒测试的所有样品中检测到特异性抗胰蛋白酶蛋白IgG抗体，但仅为市售试剂盒的四个样品。 1D-IMMUNBLOT结果表明，所有九个血清样品都是阳性的。 Spiralis ML E-S抗原，表达为特定带的存在。相比之下，血清八个样品与T. Britovi E-S ML抗原呈阳性，其中一个血清样品从33dpi（感染后的天）为阴性患者。为了鉴定由宿主抗体特异性识别的免疫反应蛋白，将两种ML E-S蛋白质与在49dPi中均采取的人血清进行2D免疫印迹。血清识别出22个蛋白斑点。螺旋和18岁。 Britovi在2D-Immunoblot分析中。它们的分子量（MW）范围为50至60kDa。 LC-MS / MS分析鉴定了常见且特异性识别的免疫反应蛋白：跨膜丝氨酸蛋白酶9，丝氨酸蛋白酶，由保护抗体靶向的抗原和肌动蛋白-1部分分别用于三氯联苯;假设蛋白质T01_7775和P49抗原，部分特异于T.螺旋; Deoxyribonuclease-2-α和假设蛋白质T03_17187 / T12_7360特异于T. Britovi。我们的结果证明了通过比较不同Trichinella物种的Xcericss的分泌物，针对寄生虫宿主关系有助于促进寄生虫宿主关系的因素的多功能工具的价值。

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