Corral M A, Paula F M, Meisel D M C L, Castilho V L P, Gonçalves E M N, Levy D, Bydlowski S P, Chieffi P P, Castro-Borges W, Gryschek R C B
Laboratório de Investigação Médica (LIM-06),Hospital das Clínicas da Faculdade de Medicina,Universidade de São Paulo,SP,Brazil.
Divisão de Laboratório Central,Hospital das Clínicas,Faculdade de Medicina,Universidade de São Paulo,São Paulo,SP,Brazil.
Parasitology. 2017 Feb;144(2):124-130. doi: 10.1017/S0031182016001645. Epub 2016 Nov 29.
Strongyloides venezuelensis is a parasitic nematode of rodents that is frequently used to obtain heterologous antigens for immunological diagnosis of human strongyloidiasis. The aim of this study was to identify antigens from filariform larvae of S. venezuelensis for immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from filariform larvae of S. venezuelensis were obtained in phosphate saline (SS and SM) and in Tris-HCl buffer (TS and TM), and were analysed by Western blotting. Different antigenic components were recognized by IgG antibodies from the sera of strongyloidiasis patients. Highest recognition was observed for a 30-40 kDa mass range present in all antigenic fractions. The band encompassing this mass range was then excised and subjected to mass spectrometry for protein identification. Immunoreactive proteins identified in the soluble fractions corresponded to metabolic enzymes, whereas cytoskeletal proteins and galectins were more abundant in the membrane fractions. These results represent the first approach towards identification of S. venezuelensis antigens for use in immunodiagnostic assays for human strongyloidiasis.
委内瑞拉粪类圆线虫是啮齿动物的一种寄生线虫,常用于获取异源抗原来进行人类类圆线虫病的免疫诊断。本研究的目的是鉴定委内瑞拉粪类圆线虫丝状蚴的抗原,用于人类类圆线虫病的免疫诊断。从委内瑞拉粪类圆线虫丝状蚴中获得的可溶性和膜部分,分别用磷酸盐缓冲液(SS和SM)和Tris-HCl缓冲液(TS和TM)提取,并通过蛋白质印迹法进行分析。类圆线虫病患者血清中的IgG抗体识别出了不同的抗原成分。在所有抗原部分中,观察到对30 - 40 kDa质量范围的识别最高。然后切下包含该质量范围的条带,进行质谱分析以鉴定蛋白质。在可溶性部分中鉴定出的免疫反应性蛋白对应于代谢酶,而细胞骨架蛋白和半乳糖凝集素在膜部分中更为丰富。这些结果代表了鉴定用于人类类圆线虫病免疫诊断检测的委内瑞拉粪类圆线虫抗原的首次尝试。