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Development and characterization of neutralizing monoclonal antibodies against canine distemper virus hemagglutinin protein

机译:抗犬瘟热病毒血凝素蛋白中和单克隆抗体的开发与表征

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摘要

Canine distemper virus (CDV) causes a serious multisystemic disease in dogs and other carnivora. Hemagglutinin (H) protein-specific antibodies are mainly responsible for protective immunity against CDV infection. In the present study, six neutralizing MAbs to the H protein of CDV were newly obtained and characterized by immunizing BALB/c mice with a recent Chinese field isolate. Competitive binding inhibition assay revealed that they recognized four distinct antigenic regions of the H protein. Immunofluorescence assay and western blotting showed that all MAbs recognize the conformational rather than the linear epitopes of the H protein. Furthermore, in immunofluorescence and virus neutralization assays, two of the MAbs were found to react only with the recent Chinese field isolate and not with older CDV strains, including vaccine strain Onderstepoort, indicating there are neutralization-related antigenic variations between the recent Chinese field isolate and the older CDV strains examined in this study. The newly established MAbs are useful for differentiating the expanding CDV strains and could be used in immunotherapy and immunodiagnosis against infection with CDV.
机译:犬瘟热病毒(CDV)在狗和其他食肉动物中引起严重的多系统疾病。血凝素(H)蛋白特异性抗体主要负责针对CDV感染的保护性免疫。在本研究中,新获得了六种针对CDV H蛋白的中和单克隆抗体,并通过用最近的中国田间分离株免疫BALB / c小鼠进行了表征。竞争性结合抑制试验表明,它们识别了H蛋白的四个不同的抗原区域。免疫荧光分析和蛋白质印迹显示,所有MAb都识别H蛋白的构象而不是线性表位。此外,在免疫荧光和病毒中和试验中,发现两个单克隆抗体仅与最近的中国田间分离株反应,而不与较老的CDV菌株(包括疫苗株Onderstepoort)反应,表明最近的中国田间分离株之间存在与中和相关的抗原变异以及本研究中检查的较老的CDV菌株。新近建立的单克隆抗体可用于区分正在扩大的CDV菌株,并可用于针对CDV感染的免疫疗法和免疫诊断。

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