Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against African Horse Sickness Virus Serotype 1 VP2 Protein.

IF 3.8 3区 医学 Q2 VIROLOGY
Viruses-Basel Pub Date : 2024-11-15 DOI:10.3390/v16111780
Xiaohua Ma, Yingzhi Zhang, Lei Na, Ting Qi, Weiwei Ma, Xing Guo, Xue-Feng Wang, Xiaojun Wang
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Abstract

African horse sickness (AHS) is an acute, fatal, contagious disease of animals of the family Equidae and is caused by infection with the African horse sickness virus (AHSV). Based on the outer capsid protein VP2, AHSV is classified into nine serotypes (AHSV-1 to -9) with little or no serological cross-reactivity between them. In 2020, AHS outbreaks caused by AHSV-1 were reported in Thailand and Malaysia, marking the first occurrences of AHS in Southeast Asia. However, little is known about the antigenic profile of AHSV-1 VP2. In this study, a recombinant VP2 protein was expressed in Escherichia coli and used as an immunogen, and three monoclonal antibodies (mAbs), designated 7D11, 10A9, and 9E7, against AHSV-1 VP2, were generated. These three mAbs were then successfully used in IFA, WB, and ELISA for the detection of AHSV-1 VP2. Two overlapping linear epitopes, 670NEFDFE675 (E670-675) recognized by 9E7 and 670NEFDF674 (E670-674) recognized by 7D11 and 10A9, were identified through truncation of GST-fused VP2. Amino acid sequence alignment shows that the 670NEFDFE675 motif is completely conserved within AHSV-1 but is highly divergent in other AHSV serotypes. Our studies provide an important tool for basic research into AHSV-1 and for the diagnosis of AHSV-1.

非洲马瘟病毒血清型 1 VP2 蛋白单克隆抗体线性表位的鉴定与特征描述
非洲马瘟(AHS)是由非洲马瘟病毒(AHSV)感染引起的马科动物的一种急性、致命的传染性疾病。根据外囊蛋白 VP2,非洲马瘟病毒可分为九个血清型(AHSV-1 至 -9),它们之间几乎没有血清学交叉反应。2020 年,泰国和马来西亚报告了由 AHSV-1 引起的 AHS 疫情爆发,这是东南亚首次出现 AHS 疫情。然而,人们对 AHSV-1 VP2 的抗原谱知之甚少。在这项研究中,在大肠杆菌中表达了重组 VP2 蛋白,并将其作为免疫原,生成了三种针对 AHSV-1 VP2 的单克隆抗体(mAbs),分别命名为 7D11、10A9 和 9E7。这三种 mAbs 被成功地用于 IFA、WB 和 ELISA 检测 AHSV-1 VP2。通过截短 GST 融合的 VP2,确定了两个重叠的线性表位,即 9E7 识别的 670NEFDFE675 (E670-675)和 7D11 及 10A9 识别的 670NEFDF674 (E670-674)。氨基酸序列比对显示,670NEFDFE675图案在AHSV-1中完全保守,但在其他AHSV血清型中却有很大差异。我们的研究为 AHSV-1 的基础研究和 AHSV-1 的诊断提供了重要工具。
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来源期刊
Viruses-Basel
Viruses-Basel VIROLOGY-
CiteScore
7.30
自引率
12.80%
发文量
2445
审稿时长
1 months
期刊介绍: Viruses (ISSN 1999-4915) is an open access journal which provides an advanced forum for studies of viruses. It publishes reviews, regular research papers, communications, conference reports and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. We also encourage the publication of timely reviews and commentaries on topics of interest to the virology community and feature highlights from the virology literature in the ''News and Views'' section. Electronic files or software regarding the full details of the calculation and experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material.
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