Immunoproteomics Identification of Major IgE and IgG4 Reactive Schistosoma japonicum Adult Worm Antigens Using Chronically Infected Human Plasma.
Immunoproteomics Identification of Major IgE and IgG4 Reactive Schistosoma japonicum Adult Worm Antigens Using Chronically Infected Human Plasma.
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DOI：
10.2149/tmh.2012-16
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发表时间：
2012-09
影响因子：
4.5
通讯作者：
Hirayama K
中科院分区：
文献类型：
--
作者：
Boamah D;Kikuchi M;Huy NT;Okamoto K;Chen H;Ayi I;Boakye DA;Bosompem KM;Hirayama K
来自流行区的免疫流行病学研究表明，年龄依赖性耐药性与血吸虫可溶性蠕虫抗原反应中IgE水平升高和IgG4抗体水平下降有关。然而，关于在血吸虫病慢性期引起IgE和IgG4免疫应答的主要抗原的分析研究有限。在本研究中，首次应用免疫蛋白质组学方法从慢性感染人群的血浆中鉴定出可被IgE和IgG4抗体识别的液体组分中的日本血吸虫抗原。ProteomeLabPF用混合的高IgE/IgG4反应性血浆样本，用斑点印迹技术筛选SWA抗原的2D分级1-D和2-D部分。在一维组分中，以较少的抗原组分检出IgE亚型(43.2%)。最常见的同工型是IgG3(79.5%)，其次是IgG1(75.0%)和IgG4(61.4%)。对反应性2-D组分进行的液相色谱-MS/MS蛋白质测序显示，18种蛋白质被鉴定、表征和确定了基因本体论类别。含锌指、RanBP2型、含结构域蛋白的2-D组分可被IgE强烈识别，而含有泛素结合酶和胞质II 5‘-核苷酸酶等蛋白质的组分可被Ig G亚类(Ig G1、Ig G3和Ig G4)强烈识别，但不能识别Ig E。本研究建立了一种简便、重复性好的鉴定日本血吸虫主要免疫活性抗原的蛋白质组学方法。预计这将促进对已确定的蛋白质的免疫原性和保护潜力的进一步研究，以及具有治疗重要性的新化合物的发现。
Immunoepidemiological studies from endemic areas have revealed age-dependent resistance correlation with increased level of IgE and decreased level of IgG4 antibodies in responses to schistosomes’ soluble worm antigen. However, there have been limited studies on analyses of major antigens that provoke IgE and IgG4 immune response during chronic stage of schistosomiasis. In this study, for the first time, immunoproteomics approach has been applied to identify S. japonicum worm antigens in liquid fractions that are recognized by IgE and IgG4 antibody using plasma from chronically infected population. ProteomeLabPF 2D fractionated 1-D and 2-D fractions of SWA antigens were screened using pooled high IgE/IgG4 reactive plasma samples by dot-blot technique. In 1-D fractions, IgE isotype was detected by fewer antigenic fractions (43.2%). The most recognized isotype was IgG3 (79.5%) followed by IgG1 (75.0%) and IgG4 (61.4%). Liquid chromatography MS/MS protein sequencing of reactive 2-D fractions revealed 18 proteins that were identified, characterized and gene ontology categories determined. 2-D fractions containing proteins such as zinc finger, RanBP2-type, domain-containing protein were strongly recognized by IgE and moderately by IgG4 whereas fractions containing proteins such as ubiquitin-conjugating enzyme and cytosolic II 5'-nucleotidase strongly recognizing by IgG subclasses (IgG1, IgG3 and IgG4) but not IgE. By this study, a simple and reproducible proteomic method has been established to identify major immunoreactive S. japonicum antigens. It is anticipated that this will stimulate further research on the immunogenicity and protective potential of proteins identified as well as discovery of novel compounds that have therapeutic importance.