Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species.
Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species.
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SARS冠状病毒尖峰和核皮质蛋白的免疫主导区域的确定和应用由不同动物物种的血清识别。
DOI：
10.1016/j.jim.2007.11.009
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发表时间：
2008-02-29
影响因子：
2.2
通讯作者：
Wang LF
中科院分区：
文献类型：
--
作者：
Yu M;Stevens V;Berry JD;Crameri G;McEachern J;Tu C;Shi Z;Liang G;Weingartl H;Cardosa J;Eaton BT;Wang LF
了解主要病毒抗原的免疫优势区对于合理设计有效的疫苗和诊断试验是很重要的。尽管有许多关于SARS-CoV的研究报告，但这些研究主要集中在人类和小鼠的免疫反应上。在这项研究中，我们的目的是寻找和比较免疫优势区的刺突蛋白（S）和核衣壳蛋白（N），这是识别不同的动物物种，包括小鼠，大鼠，兔，果子狸，猪和马的血清。在大肠杆菌中产生了12个重叠的重组蛋白片段，S和N蛋白各6个，其覆盖了两种蛋白的整个编码区。使用基于膜条的蛋白质印迹方法，确定每个抗原片段对一组动物血清的反应性。免疫显性区域含有线性表位，与所有物种的血清测试，确定了这两种蛋白质。S3片段（aa 402-622）和N4片段（aa 220-336）分别是6个S和N片段中最具免疫优势的片段。针对S3片段产生的抗体能够在ELISA中阻断一组S特异性单克隆抗体（mAb）与SARS-CoV的结合，进一步证明了该区域的免疫优势。基于这些发现，建立了一步竞争ELISA，能够检测来自人类和至少7种不同动物物种的SARS-CoV抗体。考虑到已知大量动物物种对SARS-CoV易感，这些检测将是追踪SARS-CoV起源和传播的有用工具，并将动物到人类传播的风险降至最低。
Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and diagnostic tests. Although there have been many reports of such work done for SARS–CoV, these were mainly focused on the immune responses of humans and mice. In this study, we aim to search for and compare immunodominant regions of the spike (S) and nucleocapsid (N) proteins which are recognized by sera from different animal species, including mouse, rat, rabbit, civet, pig and horse. Twelve overlapping recombinant protein fragments were produced in Escherichia coli, six each for the S and N proteins, which covered the entire coding region of the two proteins. Using a membrane-strip based Western blot approach, the reactivity of each antigen fragment against a panel of animal sera was determined. Immunodominant regions containing linear epitopes, which reacted with sera from all the species tested, were identified for both proteins. The S3 fragment (aa 402–622) and the N4 fragment (aa 220–336) were the most immunodominant among the six S and N fragments, respectively. Antibodies raised against the S3 fragment were able to block the binding of a panel of S-specific monoclonal antibodies (mAb) to SARS–CoV in ELISA, further demonstrating the immunodominance of this region. Based on these findings, one-step competition ELISAs were established which were able to detect SARS–CoV antibodies from human and at least seven different animal species. Considering that a large number of animal species are known to be susceptible to SARS–CoV, these assays will be a useful tool to trace the origin and transmission of SARS–CoV and to minimise the risk of animal-to-human transmission.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
2.2
作者：
Manopo, I;Lu, LQ;Kwang, J
通讯作者：
Kwang, J
影响因子：
11.8
作者：
Tu C;Crameri G;Kong X;Chen J;Sun Y;Yu M;Xiang H;Xia X;Liu S;Ren T;Yu Y;Eaton BT;Xuan H;Wang LF
通讯作者：
Wang LF
影响因子：
3.1
作者：
Chou, CF;Shen, S;Hong, WJ
通讯作者：
Hong, WJ
影响因子：
3.5
作者：
Wang, LF;Yu, M;Eaton, BT
通讯作者：
Eaton, BT