Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan.
Microbiol Immunol. 2009 Dec;53(12):667-74. doi: 10.1111/j.1348-0421.2009.00176.x.
The gene for phosphoprotein (P) of CDV encodes three different proteins, P, V, and C. The P protein is involved in viral gene transcription and replication. In the present study, we produced MAbs against a unique domain of the CDV-P protein, from aa 232 to 507, and determined their antigenic sites. By immunizing BALB/c mice with the recombinant P protein-specific fragment, we obtained six MAbs. Competitive binding inhibition assays revealed that they recognized two distinct regions of the P protein. Western blot analysis and immunofluorescence assays using deletion mutants of the unique C-terminus of the CDV-P protein revealed that all MAbs recognized a central short region (aa 233-303) of the CDV-P protein. In addition, linear and conformational epitopes have been determined, and at least four antigenic sites exist in the P protein central region. Furthermore, four of the MAbs were found to react with the P protein of recent Japanese field isolates but not with that of the older CDV strains, including a vaccine strain. Thus, these MAbs could be clinically useful for quick diagnosis during the CDV outbreaks.
犬瘟热病毒（CDV）磷蛋白（P）基因编码三种不同的蛋白，即 P、V 和 C。P 蛋白参与病毒基因转录和复制。本研究中，我们制备了针对 CDV-P 蛋白独特结构域（aa232-507）的单克隆抗体（MAbs），并鉴定了其抗原表位。通过用重组 P 蛋白特异性片段免疫 BALB/c 小鼠，我们获得了 6 株 MAb。竞争性结合抑制试验表明，它们识别 P 蛋白的两个不同区域。Western blot 分析和用 CDV-P 蛋白独特 C 末端缺失突变体进行的免疫荧光试验表明，所有 MAb 均识别 CDV-P 蛋白的中央短区域（aa233-303）。此外，已确定线性和构象表位，并且在 P 蛋白中央区域存在至少四个抗原表位。此外，发现其中 4 株 MAb 与近期日本田间分离株的 P 蛋白反应，但与较旧的 CDV 株（包括疫苗株）不反应。因此，这些 MAb 可能在 CDV 爆发期间的快速诊断中具有临床应用价值。
