Sotillo Javier, Sanchez-Flores Alejandro, Cantacessi Cinzia, Harcus Yvonne, Pickering Darren, Bouchery Tiffany, Camberis Mali, Tang Shiau-Choot, Giacomin Paul, Mulvenna Jason, Mitreva Makedonka, Berriman Matthew, LeGros Graham, Maizels Rick M, Loukas Alex
From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
§Parasite Genomics, Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK;
Mol Cell Proteomics. 2014 Oct;13(10):2736-51. doi: 10.1074/mcp.M114.038950. Epub 2014 Jul 3.
Hookworms infect more than 700 million people worldwide and cause more morbidity than most other human parasitic infections. Nippostrongylus brasiliensis (the rat hookworm) has been used as an experimental model for human hookworm because of its similar life cycle and ease of maintenance in laboratory rodents. Adult N. brasiliensis, like the human hookworm, lives in the intestine of the host and releases excretory/secretory products (ESP), which represent the major host-parasite interface. We performed a comparative proteomic analysis of infective larval (L3) and adult worm stages of N. brasiliensis to gain insights into the molecular bases of host-parasite relationships and determine whether N. brasiliensis could indeed serve as an appropriate model for studying human hookworm infections. Proteomic data were matched to a transcriptomic database assembled from 245,874,892 Illumina reads from different developmental stages (eggs, L3, L4, and adult) of N. brasiliensis yielding∼18,426 unigenes with 39,063 possible isoform transcripts. From this analysis, 313 proteins were identified from ESPs by LC-MS/MS-52 in the L3 and 261 in the adult worm. Most of the proteins identified in the study were stage-specific (only 13 proteins were shared by both stages); in particular, two families of proteins-astacin metalloproteases and CAP-domain containing SCP/TAPS-were highly represented in both L3 and adult ESP. These protein families are present in most nematode groups, and where studied, appear to play roles in larval migration and evasion of the host's immune response. Phylogenetic analyses of defined protein families and global gene similarity analyses showed that N. brasiliensis has a greater degree of conservation with human hookworm than other model nematodes examined. These findings validate the use of N. brasiliensis as a suitable parasite for the study of human hookworm infections in a tractable animal model.
钩虫感染了全球超过7亿人,其导致的发病率高于大多数其他人体寄生虫感染。巴西日圆线虫(大鼠钩虫)因其与人类钩虫相似的生命周期以及在实验室啮齿动物中易于饲养,已被用作人类钩虫的实验模型。与人类钩虫一样,成年巴西日圆线虫生活在宿主的肠道中,并释放排泄/分泌产物(ESP),这是宿主与寄生虫的主要界面。我们对巴西日圆线虫的感染性幼虫(L3)和成虫阶段进行了比较蛋白质组学分析,以深入了解宿主 - 寄生虫关系的分子基础,并确定巴西日圆线虫是否确实可以作为研究人类钩虫感染的合适模型。蛋白质组学数据与一个转录组数据库进行匹配,该转录组数据库由来自巴西日圆线虫不同发育阶段(卵、L3、L4和成虫)的245,874,892条Illumina读数组装而成,产生了约18,426个单基因,具有39,063个可能的异构体转录本。通过该分析,通过LC-MS/MS从ESP中鉴定出313种蛋白质 - L3阶段有52种,成虫阶段有261种。研究中鉴定出的大多数蛋白质是阶段特异性的(两个阶段仅共有13种蛋白质);特别是,两个蛋白质家族 - 阿斯他汀金属蛋白酶和含CAP结构域的SCP/TAPS - 在L3和成虫ESP中都有高度表达。这些蛋白质家族存在于大多数线虫类群中,并且在已研究的情况下,似乎在幼虫迁移和逃避宿主免疫反应中发挥作用。对特定蛋白质家族的系统发育分析和全局基因相似性分析表明,与其他所研究的模型线虫相比,巴西日圆线虫与人类钩虫具有更高程度的保守性。这些发现证实了巴西日圆线虫作为一种合适的寄生虫,可用于在易于处理的动物模型中研究人类钩虫感染。