Laboratório de Investigação Médica (LIM/06), Hospital das Clinicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
Instituto de Medicina Tropical, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
Parasitol Res. 2022 Nov;121(11):3155-3170. doi: 10.1007/s00436-022-07636-y. Epub 2022 Aug 31.
Serodiagnosis of human strongyloidiasis is a practical alternative to parasitological methods due to its high sensitivity. However, cross-reactivity with other helminth infections limits its utility, and this problem is due to the use of homologous or heterologous somatic extracts of the parasite as an antigen source. Excretory-secretory (E/S) products from Strongyloides infective larvae can be used to improve the serodiagnosis. The combined use of western blot and proteomics became an interesting strategy to identify immunological markers for the serodiagnosis of strongyloidiasis. The present study describes the proteomic analysis of the antigenic components from E/S products of S. venezuelensis infective larvae that were recognized by IgG antibodies from patients with strongyloidiasis. Our results showed that IgG antibodies from patients with strongyloidiasis recognized between 15 and 16 antigenic bands in the E/S products from S. venezuelensis that were incubated in PBS or in RPMI culture medium, respectively. Overall, antigenic bands of low and high molecular weight were more specific than those of intermediate molecular weight, which were cross-reactive. A 36-kDa antigenic band was 93% sensitive and 100% specific (a probably arginine kinase of 37 kDa), while other antigenic bands were highly sensitive but low specific. Proteomic analysis revealed differences between the protein profiles from E/S-RPMI and E/S-PBS since only one-third of all proteins identified were common in both types of E/S products. Bioinformatic analysis showed that more than 50% of the proteins from E/S products are secreted within extracellular vesicles and only a small percentage of them are actually released by the classical secretory pathway. Several components from the E/S products were identified as plasminogen-binding proteins, probably used as an immune evasion mechanism. The data provided here provide valuable information to increase understanding of E/S products from S. venezuelensis infective larvae. This may help us to find new targets for the immunodiagnosis of human strongyloidiasis.
人旋毛虫病的血清学诊断是寄生虫学方法的实用替代方法,因为它具有很高的敏感性。然而,与其他寄生虫感染的交叉反应限制了其用途,而这个问题是由于使用寄生虫的同源或异源体提取物作为抗原来源。旋毛虫感染幼虫的排泄-分泌(E/S)产物可用于改善血清学诊断。Western blot 和蛋白质组学的联合使用成为鉴定旋毛虫病血清学诊断免疫标志物的一种有趣策略。本研究描述了旋毛虫 E/S 产物中抗原成分的蛋白质组学分析,这些成分被旋毛虫病患者的 IgG 抗体识别。我们的结果表明,旋毛虫病患者的 IgG 抗体在 PBS 或 RPMI 培养基中孵育的 S. venezuelensis E/S 产物中识别了 15 到 16 个抗原带。总体而言,低和高分子量的抗原带比中分子量的抗原带更具特异性,而中分子量的抗原带则具有交叉反应性。36 kDa 的抗原带具有 93%的敏感性和 100%的特异性(可能是 37 kDa 的精氨酸激酶),而其他抗原带则具有高敏感性但低特异性。蛋白质组学分析显示 E/S-RPMI 和 E/S-PBS 之间的蛋白质图谱存在差异,因为在两种 E/S 产物中鉴定出的所有蛋白质中只有三分之一是共同的。生物信息学分析表明,E/S 产物中的蛋白质有超过 50%是通过细胞外囊泡分泌的,只有一小部分是通过经典的分泌途径释放的。E/S 产物中的几种成分被鉴定为纤溶酶原结合蛋白,可能被用作免疫逃避机制。这里提供的数据为增加对 S. venezuelensis 感染幼虫 E/S 产物的理解提供了有价值的信息。这可能有助于我们找到人旋毛虫病免疫诊断的新靶点。