从巨型片形吸虫中发现的一种新型硫氧还蛋白相关蛋白 14 具有用于片形吸虫病的免疫诊断潜力。
A novel Thioredoxin-related protein 14 from Fasciola gigantica has an immunodiagnostic potential for fasciolosis.
机构信息
Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.; Research unit of vaccine and diagnosis of parasitic diseases, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.
Division of Anatomy, Department of Preclinical Science, Faculty of Medicine, Thammasat University, Pathumthani 12120, Thailand; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand.
出版信息
Acta Trop. 2020 Jul;207:105471. doi: 10.1016/j.actatropica.2020.105471. Epub 2020 Apr 14.
In the definitive host, a trematode parasite can survive and evade the damage by reactive oxygen species that are generated from its metabolism and the host immune cells. Several anti-oxidant proteins are found in Fasciola spp. which play essential roles in cellular redox balance. One of them is thioredoxin-related protein 14 (TRP14) that has a highly conserved WCPDC motif and serves as a disulfide reductase-like thioredoxin (Trx). In the present study, a cDNA encoding TRP14 from F. gigantica (FgTRP14) was selected and cloned by immunoscreening with a rabbit infected serum. Phylogenetic analysis was performed by MEGA X program showed that FgTRP14 was most highly related to the Fasciola hepatica. Immunoblotting analysis of the polyclonal antibody rabbit serum against recombinant FgTRP14 (rFgTRP14) revealed that the molecular weight of natural FgTRP14 was at 14 kDa from metacercariae, NEJ, 4-week old juvenile and adult stage. The native FgTRP14 was expressed in caecal epithelial cells and preferentially localized on the cells' surface lamellae of adult stage. By sandwich ELISA assay, the circulating FgTRP14 could be recognized in sera of experimentally F. gigantica metacercariae infection in mice. The native FgTRP14 in the excretory-secretory (ES) and whole body (WB) of adult F. gigantica were detected at the concentrations 6.3 ng/ml, and 45 ng/ml, respectively. Therefore, it could be considered for immunodiagnostic candidate for fasciolosis.
在终末宿主中，肝片形吸虫寄生虫可以存活并逃避其代谢和宿主免疫细胞产生的活性氧物质的损害。在双腔吸虫属中发现了几种抗氧化蛋白，它们在细胞氧化还原平衡中发挥重要作用。其中之一是硫氧还蛋白相关蛋白 14（TRP14），它具有高度保守的 WCPDC 基序，是一种类似二硫键还原酶的硫氧还蛋白（Trx）。本研究通过用感染兔血清免疫筛选，从 F. gigantica 中选择并克隆了编码 TRP14 的 cDNA（FgTRP14）。通过 MEGA X 程序进行的系统发育分析表明，FgTRP14 与 Fasciola hepatica 最为密切相关。针对重组 FgTRP14（rFgTRP14）的多克隆兔血清的免疫印迹分析表明，天然 FgTRP14 的分子量在囊蚴、NEJ、4 周龄幼体和成体阶段均为 14 kDa。天然 FgTRP14 在盲肠上皮细胞中表达，并优先定位于成体阶段细胞的表面薄片上。通过夹心 ELISA 测定，可在感染肝片形吸虫囊蚴的实验小鼠血清中检测到循环 FgTRP14。在成年 F. gigantica 的排泄-分泌（ES）和全身（WB）中分别检测到天然 FgTRP14 的浓度为 6.3 ng/ml 和 45 ng/ml。因此，它可以被认为是肝片形吸虫病的免疫诊断候选物。
Zotero 插件