Xue W, Minocha H C
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506, USA.
Vet Microbiol. 1996 Mar;49(1-2):67-79. doi: 10.1016/0378-1135(95)00167-0.
Anti-idiotypic antibodies (anti-ids) have been used successfully in studies on bovine viral diarrhea virus (BVDV) receptor(s) in our laboratory. The anti-ids specifically bound to cultured cells and identified a 50 kDa cellular membrane protein, which is thought to be a specific receptor for BVDV. In this study, flow cytometric analyses demonstrated that the anti-ids also specifically bound to different cell types, namely MDBK, EBK, BT, PK15, MA1O4, and Vero. Experiments on virus attachment and replication showed that BVDV adsorbed to all cells and replicated in them except monkey kidney cells MA 104 and Vero (non-permissive). Results from plaque reduction assays indicated that cellular membrane proteins from all cell lines competitively inhibited BVDV attachment to cultured MDBK cells, suggesting the presence of BVDV receptor on all cells. Immunoblotting of cell membrane proteins with the anti-ids revealed a 50 kDa protein in both permissive and nonpermissive cells. Subcloned or synchronized MDBK cells demonstrated no significant difference of binding with anti-ids as compared to normal cultured cells.
抗独特型抗体(抗-ids)已在我们实验室成功用于牛病毒性腹泻病毒(BVDV)受体的研究。抗-ids与培养细胞特异性结合,并鉴定出一种50 kDa的细胞膜蛋白,该蛋白被认为是BVDV的特异性受体。在本研究中,流式细胞术分析表明,抗-ids也与不同细胞类型特异性结合,即MDBK、EBK、BT、PK15、MA1O4和Vero细胞。病毒吸附和复制实验表明,BVDV能吸附到所有细胞并在其中复制,但猴肾细胞MA 104和Vero细胞(非允许细胞)除外。蚀斑减少试验结果表明,所有细胞系的细胞膜蛋白均能竞争性抑制BVDV对培养的MDBK细胞的吸附,提示所有细胞上均存在BVDV受体。用抗-ids对细胞膜蛋白进行免疫印迹分析显示,允许细胞和非允许细胞中均存在一种50 kDa的蛋白。亚克隆或同步化的MDBK细胞与正常培养细胞相比,与抗-ids的结合无显著差异。