Antibody profiles to wheat germ cell-free system synthesized Plasmodium falciparum proteins correlate with protection from symptomatic malaria in Uganda
Antibody profiles to wheat germ cell-free system synthesized Plasmodium falciparum proteins correlate with protection from symptomatic malaria in Uganda
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DOI：
10.1016/j.vaccine.2017.01.001
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发表时间：
2017-02-07
期刊：
影响因子：
5.5
通讯作者：
Tsuboi, Takafumi
中科院分区：
文献类型：
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作者：
Kanoi, Bernard N.;Takashima, Eizo;Tsuboi, Takafumi
针对恶性疟原虫的保护性抗体的关键目标在很大程度上仍然未知。在这项研究中，我们确定了1827个重组蛋白的免疫反应性，这些重组蛋白来自1565个基因，占整个恶性疟原虫基因组的30%，用于鉴定新型疟疾疫苗候选物。利用小麦胚无细胞系统表达重组蛋白，该系统是一个可以合成高质量疟原虫蛋白的平台，可以在动物体内诱导具有生物活性的抗体。血清取自乌干达北方疟疾流行地区的土著居民，他们在雨季开始时被招募，并前瞻性监测一年的疟疾症状。通过AlphaScreen测定对血清的免疫反应性; AlphaScreen是一种检测蛋白质相互作用的均相高通量系统。我们的分析揭示了对128种蛋白质的抗体应答，这些蛋白质与预防症状性疟疾显著相关。从128种蛋白质中，53种由于具有预测的信号肽和/或跨膜结构域或确认在寄生虫表面上的定位而被向下选择为宿主保护性免疫应答的最合理的靶标。这53种蛋白质不仅包括先前表征的候选疫苗，而且包括未表征的蛋白质。参与红细胞侵袭的蛋白质; RON 4、RON 2和CLAG3.1以及红细胞前蛋白质; SIAP-2、TRAP和CelTOS被推荐作为优先考虑的候选疫苗进一步评价。这些发现清楚地表明，使用小麦胚芽无细胞系统结合AlphaScreen高通量免疫筛选产生蛋白质文库为合理发现和选择潜在的疟疾候选疫苗提供了新的选择。(C)2017爱思唯尔有限公司版权所有。
The key targets of protective antibodies against Plasmodium falciparum remain largely unknown. In this study, we determined immunoreactivity to 1827 recombinant proteins derived from 1565 genes representing similar to 30% of the entire P. falciparum genome, for identification of novel malaria vaccine candidates. The recombinant proteins were expressed by wheat germ cell-free system, a platform that can synthesize quality plasmodial proteins that elicit biologically active antibodies in animals. Sera were obtained from indigenous residents of a malaria endemic region in Northern Uganda who were enrolled at the start of a rainy season and prospectively monitored for symptomatic malaria episodes for a year. Immunoreactivity to sera was determined by AlphaScreen; a homogeneous high-throughput system that detects protein interactions. Our analysis revealed antibody responses to 128 proteins that significantly associated with protection from symptomatic malaria. From 128 proteins, 53 were down-selected as the most plausible targets of host protective immune response by virtue of having a predicted signal peptide and/or trans membrane domain(s), or confirmed localization on the parasite surface. The 53 proteins comprised of not only previously characterized vaccine candidates but also uncharacterized proteins. Proteins involved in erythrocyte invasion; RON4, RON2 and CLAG3.1 and pre-erythrocytic proteins; SIAP-2, TRAP and CelTOS, were recommended for prioritization for further evaluation as vaccine candidates. The findings clearly demonstrate that generation of the protein library using the wheat germ cell-free system coupled with high throughput immunoscreening with AlphaScreen offers new options for rational discovery and selection of potential malaria vaccine candidates. (C) 2017 Elsevier Ltd. All rights reserved.