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Identification of porcine alveolar macrophage glycoproteins involved in infection of porcine respiratory and reproductive syndrome virus.

机译:鉴定涉及猪呼吸道和生殖综合症病毒感染的猪肺泡巨噬细胞糖蛋白。

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摘要

The aim of this study was to identify the receptor(s) for PRRSV on porcine alveolar macrophages (PAMs) by producing monoclonal antibodies (MAbs) against these cells. Hybridoma supernatants were selected for their ability to block PRRSV infection. Four MAbs, 1-8D2, 9.4C7, 9.9F2, and 3-3H2 inhibited infection and recognised cell surface, PAM-specific antigens as shown by immunofluorescence and immunoperoxidase monolayer assay. These MAbs were then used to identify cellular proteins involved in PRRSV infection by radioimmunoprecipitation assays (RIPAs). MAbs 1-8D2 and 9.9F2 each recognised a 150 kDa-polypeptide doublet, while MAbs 9.4C7 and 3-3H2 both recognised a 220 kDa-polypeptide. Glycosidase treatment demonstrated all these polypeptides to be N-glycosylated. Thus, multiple glycoproteins appear to be involved in infection of PAMs by PRRSV.
机译:这项研究的目的是通过产生针对这些细胞的单克隆抗体(MAb)来鉴定猪肺泡巨噬细胞(PAM)上PRRSV的受体。选择杂交瘤上清液以阻断PRRSV感染。如免疫荧光和免疫过氧化物酶单层测定所示,四种单克隆抗体1-8D2、9.4C7、9.9F2和3-3H2可抑制感染并识别细胞表面,PAM特异性抗原。然后,通过放射免疫沉淀测定法(RIPA)将这些单克隆抗体用于鉴定参与PRRSV感染的细胞蛋白。 MAb 1-8D2和9.9F2各自识别150 kDa的多肽双峰,而MAb 9.4C7和3-3H2都识别220 kDa的多肽。糖苷酶处理证明所有这些多肽都是N-糖基化的。因此,多种糖蛋白似乎与PRRSV感染PAM有关。

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