Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, 16802, USA.
Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA.
Commun Biol. 2023 Sep 19;6(1):955. doi: 10.1038/s42003-023-05319-7.
Canine parvovirus (CPV) is an important pathogen that emerged by cross-species transmission to cause severe disease in dogs. To understand the host immune response to vaccination, sera from dogs immunized with parvovirus are obtained, the polyclonal antibodies are purified and used to solve the high resolution cryo EM structures of the polyclonal Fab-virus complexes. We use a custom software, Icosahedral Subparticle Extraction and Correlated Classification (ISECC) to perform subparticle analysis and reconstruct polyclonal Fab-virus complexes from two different dogs eight and twelve weeks post vaccination. In the resulting polyclonal Fab-virus complexes there are a total of five distinct Fabs identified. In both cases, any of the five antibodies identified would interfere with receptor binding. This polyclonal mapping approach identifies a specific, limited immune response to the live vaccine virus and allows us to investigate the binding of multiple different antibodies or ligands to virus capsids.
犬细小病毒(CPV)是一种重要的病原体,通过跨物种传播而出现,可导致犬只严重疾病。为了了解宿主对疫苗接种的免疫反应,我们从接种细小病毒的犬只中获得血清,纯化多克隆抗体,并用于解决多克隆 Fab-病毒复合物的高分辨率 cryo EM 结构。我们使用自定义软件 Icosahedral Subparticle Extraction and Correlated Classification(ISECC)来进行亚颗粒分析,并从接种后 8 周和 12 周的两只不同犬只中重建多克隆 Fab-病毒复合物。在产生的多克隆 Fab-病毒复合物中总共鉴定出了 5 种不同的 Fab。在这两种情况下,鉴定出的 5 种抗体中的任何一种都可能干扰受体结合。这种多克隆定位方法确定了对活疫苗病毒的特异性、有限的免疫反应,并使我们能够研究多种不同抗体或配体与病毒衣壳的结合。