McWilliam Hamish E G, Driguez Patrick, Piedrafita David, McManus Donald P, Meeusen Els N T
Department of Microbiology and Immunology, The University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
Parasit Vectors. 2014 Jun 25;7:290. doi: 10.1186/1756-3305-7-290.
Novel vaccine candidates against Schistosoma japonicum are required, and antigens present in the vulnerable larval developmental stage are attractive targets. Post-genomic technologies are now available which can contribute to such antigen discovery.
A schistosome-specific protein microarray was probed using the local antibody response against migrating larvae. Antigens were assessed for their novelty and predicted larval expression and host-exposed features. One antigen was further characterised and its sequence and structure were analysed in silico. Real-time polymerase chain reaction was used to analyse transcript expression throughout development, and immunoblotting and enzyme-linked immunosorbent assays employed to determine antigen recognition by antibody samples.
Several known and novel antigens were discovered, two of which showed up-regulated transcription in schistosomula. One novel antigen, termed S. japonicum Ly-6-like protein 1 (Sj-L6L-1), was further characterised and shown to share structural and sequence features with the Ly-6 protein family. It was found to be present in the worm tegument and expressed in both the larval and adult worms, but was found to be antigenic only in the lungs that the larvae migrate to and traverse.
This study represents a novel approach to vaccine antigen discovery and may contribute to schistosome vaccine development against this important group of human and veterinary pathogens.
需要新型日本血吸虫疫苗候选物,而处于易损幼虫发育阶段的抗原是有吸引力的靶点。后基因组技术现已可用,可有助于此类抗原的发现。
使用针对移行幼虫的局部抗体反应探测血吸虫特异性蛋白质微阵列。评估抗原的新颖性、预测的幼虫表达和宿主暴露特征。对一种抗原进行进一步表征,并在计算机上分析其序列和结构。使用实时聚合酶链反应分析整个发育过程中的转录本表达,并采用免疫印迹和酶联免疫吸附测定来确定抗体样品对抗原的识别。
发现了几种已知和新型抗原,其中两种在童虫中显示转录上调。一种新型抗原,称为日本血吸虫Ly-6样蛋白1(Sj-L6L-1),进行了进一步表征,显示与Ly-6蛋白家族具有结构和序列特征。发现它存在于虫体被膜中,在幼虫和成虫中均有表达,但仅在幼虫迁移并穿过的肺部具有抗原性。
本研究代表了一种疫苗抗原发现的新方法,可能有助于针对这一重要的人类和兽类病原体开发血吸虫疫苗。