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首页> 外文期刊>Virology >Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV).
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Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV).

机译：通过重症急性呼吸综合征相关冠状病毒（SARS-CoV）的灭活的中国分离株免疫小鼠而产生的抗体的抗原决定部位定位和生物学功能分析。

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Inactivated severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been tested as a candidate vaccine against the re-emergence of SARS. In order to understand the efficacy and safety of this approach, it is important to know the antibody specificities generated with inactivated SARS-CoV. In the current study, a panel of twelve monoclonal antibodies (mAbs) was established by immunizing Balb/c mice with the inactivated BJ01 strain of SARS-CoV isolated from the lung tissue of a SARS-infected Chinese patient. These mAbs could recognize SARS-CoV-infected cells by immunofluorescence analysis (IFA). Seven of them were mapped to the specific segments of recombinant spike (S) protein: six on S1 subunit (aa 12-798) and one on S2 subunit (aa 797-1192). High neutralizing titers against SARS-CoV were detected with two mAbs (1A5 and 2C5) targeting at a subdomain of S protein (aa 310-535), consistent with the previous report that this segment of S protein contains the major neutralizing domain. Some of these S-specific mAbs were able to recognize cleaved products of S protein in SARS-CoV-infected Vero E6 cells. None of the remaining five mAbs could recognize either of the recombinant S, N, M, or E antigens by ELISA. This study demonstrated that the inactivated SARS-CoV was able to preserve the immunogenicity of S protein including its major neutralizing domain. The relative ease with which these mAbs were generated against SARS-CoV virions further supports that subunit vaccination with S constructs may also be able to protect animals and perhaps humans. It is somewhat unexpected that no N-specific mAbs were identified albeit anti-N IgG was easily identified in SARS-CoV-infected patients. The availability of this panel of mAbs also provided potentially useful agents with applications in therapy, diagnosis, and basic research of SARS-CoV.
机译：灭活的严重急性呼吸综合征相关冠状病毒（SARS-CoV）已作为抗SARS复发的候选疫苗进行了测试。为了了解这种方法的有效性和安全性，重要的是要了解灭活SARS-CoV产生的抗体特异性。在本研究中，通过用从SARS感染中国患者的肺组织中分离出的灭活SARS-CoV BJ01株对Balb / c小鼠进行免疫，建立了一组十二种单克隆抗体（mAb）。这些单克隆抗体可以通过免疫荧光分析（IFA）识别被SARS-CoV感染的细胞。其中七个被定位到重组刺突（S）蛋白的特定片段上：六个位于S1亚基上（aa 12-798），一个位于S2亚基上（aa 797-1192）。用针对S蛋白亚结构域（aa 310-535）的两个mAb（1A5和2C5）检测到了针对SARS-CoV的高中和效价，这与先前报道S蛋白的这一片段包含主要中和结构域一致。这些S特异性mAb中的一些能够识别SARS-CoV感染的Vero E6细胞中S蛋白的裂解产物。其余五个mAb均不能通过ELISA识别重组S，N，M或E抗原。这项研究表明，灭活的SARS-CoV能够保留S蛋白的免疫原性，包括其主要的中和结构域。这些抗SARS-CoV病毒粒子的单克隆抗体相对容易产生，这进一步证明了用S构建体进行亚基疫苗接种也可能能够保护动物甚至人类。出乎意料的是，尽管在SARS-CoV感染的患者中很容易鉴定出抗N IgG，但未鉴定出N特异性mAb。该单克隆抗体的可用性还为在SARS-CoV的治疗，诊断和基础研究中的应用提供了潜在有用的药物。

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