Profiling the humoral immune responses to Plasmodium vivax infection and identification of candidate immunogenic rhoptry-associated membrane antigen (RAMA)
Profiling the humoral immune responses to Plasmodium vivax infection and identification of candidate immunogenic rhoptry-associated membrane antigen (RAMA)
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DOI：
10.1016/j.jprot.2014.02.029
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发表时间：
2014-05-06
影响因子：
3.3
通讯作者：
Han, Eun-Taek
中科院分区：
文献类型：
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作者：
Lu, Feng;Li, Jian;Han, Eun-Taek
间日疟原虫基因组和转录组测序的完成为在> 5000种候选蛋白中鉴定抗原提供了机会。为了鉴定那些具有免疫原性的间日疟原虫蛋白，使用小麦胚芽无细胞系统表达了总共152种候选蛋白（160个片段）。Western blot分析结果显示，表达率为92.5%（148/160），其中96.6%（143/148）为可溶性表达，可溶性表达率为67.7%。利用蛋白芯片技术对22例间日疟患者和10例健康人血清进行免疫学分析，共鉴定出44个（27.5%，44/160）间日疟高反应性抗原。总体而言，5种候选物（棒状体相关膜抗原[拉马]、Pv-fam-a和-B、EXP-1和假设蛋白PVX_084775）显示阳性反应，患者血清> 80%，21种候选物50%至80%。超过23%的高免疫反应性蛋白质是假设的蛋白质，在这项研究中首次描述。最高免疫原性蛋白之一拉马被表征并证实是近期暴露于间日疟原虫感染的血清学标志物。这些新的免疫蛋白质组应大大促进有前途的新的疟疾抗原的鉴定，并可能值得进一步study.Biological significanceThe高通量克隆和表达系统的建立允许构建蛋白质阵列的蛋白质组范围内的研究间日疟原虫。在这项研究中，高通量筛选方法已被应用于调查血液阶段特异性免疫蛋白质组从间日疟原虫。我们从152个假定的候选者中鉴定了44个抗原蛋白，其中超过23%是本研究中首次描述的假设蛋白。此外，进一步表征PvRAMA，并证实其为暴露于感染的血清学标志物。三分之一的选择的抗原基因的表达之间的转移间日疟原虫和恶性疟原虫，这表明这些基因可能代表与间日疟原虫的选择性年轻的红细胞和/或与免疫逃避的重要因素。这些新的免疫蛋白质组的间日疟原虫血液阶段提供了一个基线，为进一步的前瞻性血清学标记物研究疟疾。这些方法可用于从间日疟原虫基因组中确定免疫显性候选抗原。(C)2014爱思唯尔有限公司版权所有。
Completion of sequencing of the Plasmodium vivax genome and transcriptome offers the chance to identify antigens among > 5000 candidate proteins. To identify those P. vivax proteins that are immunogenic, a total of 152 candidate proteins (160 fragments) were expressed using a wheat germ cell-free system. The results of Western blot analysis showed that 92.5% (148/160) of the targets were expressed, and 96.6% (143/148) were in a soluble form with 67.7% of solubility rate. The proteins were screened by protein arrays with sera from 22 vivax malaria patients and 10 healthy individuals to confirm their immune profile, and 44 (27.5%, 44/160) highly reactive P. vivax antigens were identified. Overall, 5 candidates (rhoptry-associated membrane antigen [RAMA], Pv-fam-a and -b, EXP-1 and hypothetical protein PVX_084775) showed a positive reaction with > 80% of patient sera, and 21 candidates with 50% to 80%. More than 23% of the highly immunoreactive proteins were hypothetical proteins, described for the first time in this study. One of the top immunogenic proteins, RAMA, was characterized and confirmed to be a serological marker of recent exposure to P. vivax infection. These novel immunoproteomes should greatly facilitate the identification of promising novel malaria antigens and may warrant further study.Biological significanceThe establishment of high-throughput cloning and expression systems has permitted the construction of protein arrays for proteome-wide study of Plasmodium vivax. In this study,high-throughput screening assays have been applied to investigate blood stage-specific immune proteomes from P. vivax. We identified 44 antigenic proteins from the 152 putative candidates, more than 23% of which were hypothetical proteins described for the first time in this study. In addition, PvRAMA was characterized further and confirmed to be a serological marker of exposure to infections. The expression of one-third of the selected antigenic genes were shifted between P. vivax and Plasmodium falciparum, suggesting that these genes may represent important factors associated with P. vivax selectivity for young erythrocytes and/or with immune evasion. These novel immune proteomes of the P. vivax blood stage provide a baseline for further prospective serological marker studies in malaria. These methods could be used to determine immunodominant candidate antigens from the P. vivax genome. (C) 2014 Elsevier B.V. All rights reserved.