Laboratório de Biologia Parasitária da Faculdade de Biociências e Laboratório de Parasitologia Molecular do Instituto de Pesquisas Biomédicas da Pontifícia Universidade do Rio Grande do Sul, Avenida Ipiranga 6690, 90690-900 Porto Alegre RS, Brazil.
Exp Parasitol. 2012 Jan;130(1):26-31. doi: 10.1016/j.exppara.2011.10.003. Epub 2011 Oct 10.
Angiostrongyliasis results from infections with intra-arterial nematodes that accidentally infect humans. Specifically, infections with Angiostrongylus cantonensis cause eosinophilic meningitis and Angiostrongylus costaricensis infections result in eosinophilic enteritis. Immunological tests are the primary means of diagnosing infections with either pathogen since these parasites are usually not recoverable in fecal or cerebrospinal fluid. However, well-defined, purified antigens are not currently available in sufficient quantities from either pathogen for use in routine immunodiagnostic assays. Since A. costaricensis and A. cantonensis share common antigens, sera from infected persons will recognize antigens from either species. In addition to their potential use in angiostrongyliasis diagnosis, characterization of these proteins that establish the host-parasite interphase would improve our understanding of the biology of these parasites. The main objective of the present work was to characterize A. cantonensis excretory-secretory (ES) products by analyzing ES preparations by two-dimensional gel electrophoresis coupled with immunoblotting using pools of positive sera (PS) and sera from healthy individuals (SC). Protein spots recognized by PS were excised and analyzed by electrospray ionization (ESI) mass spectrometry. MASCOT analysis of mass spectrometry data identified 17 proteins: aldolase; CBR-PYP-1 protein; beta-amylase; heat shock protein 70; proteosome subunit beta type-1; actin A3; peroxiredoxin; serine carboxypeptidase; protein disulfide isomerase 1; fructose-bisphosphate aldolase 2; aspartyl protease inhibitor; lectin-5; hypothetical protein F01F1.12; cathepsin B-like cysteine proteinase 1; hemoglobinase-type cysteine proteinase; putative ferritin protein 2; and a hypothetical protein. Molecular cloning of these respective targets will next be carried out to develop a panel of Angiostrongylus antigens that can be used for diagnostic purposes and to further study host-Angiostrongylus interactions.
血管圆线虫病是由偶然感染人体的动脉内线虫引起的。具体来说，感染广州管圆线虫会导致嗜酸性脑膜炎，而感染食酸性旋毛虫则会导致嗜酸性肠炎。由于这些寄生虫通常在粪便或脑脊液中无法恢复，因此免疫测试是诊断这两种病原体感染的主要手段。然而，由于这两种病原体都没有足够数量的明确、纯化的抗原可用于常规免疫诊断检测，因此目前无法使用。由于 A. costaricensis 和 A. cantonensis 具有共同的抗原，因此感染患者的血清会识别来自任何一种物种的抗原。除了它们在血管圆线虫病诊断中的潜在用途外，这些建立宿主-寄生虫相互作用的蛋白质的特征描述将有助于我们更好地了解这些寄生虫的生物学特性。本研究的主要目的是通过二维凝胶电泳结合使用阳性血清（PS）和健康个体血清（SC）池进行免疫印迹分析，来分析 A. cantonensis 排泄分泌（ES）产物。使用 PS 识别的蛋白质斑点被切除并通过电喷雾电离（ESI）质谱进行分析。质谱数据分析的 MASCOT 分析鉴定了 17 种蛋白质：醛缩酶；CBR-PYP-1 蛋白；β-淀粉酶；热休克蛋白 70；蛋白酶体亚基β型-1；肌动蛋白 A3；过氧化物酶；丝氨酸羧肽酶；蛋白质二硫键异构酶 1；果糖-二磷酸醛缩酶 2；天冬氨酸蛋白酶抑制剂；凝集素-5；假设蛋白 F01F1.12；组织蛋白酶 B 样半胱氨酸蛋白酶 1；血红蛋白酶型半胱氨酸蛋白酶；假定铁蛋白蛋白 2；和一个假设蛋白。接下来将对这些靶标进行分子克隆，以开发一组可用于诊断目的的血管圆线虫抗原，并进一步研究宿主-血管圆线虫相互作用。
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