Barnett P V, Samuel A R, Pullen L, Ansell D, Butcher R N, Parkhouse R M
Institute for Animal Health, Pirbright Laboratory, Surrey, UK.
J Gen Virol. 1998 Jul;79 ( Pt 7):1687-97. doi: 10.1099/0022-1317-79-7-1687.
Eight neutralizing and two non-neutralizing anti-foot-and-mouth disease virus (FMDV) bovine IgG1 and IgG2 monoclonal antibodies (BMAbs) recognize conformationally dependent epitopes. The majority of those shown to neutralize virus passively protected mice from virus challenge, regardless of isotype. Well-characterized anti-FMDV mouse MAbs, representing five independent neutralizing epitopes on O1 serotype virus, were examined with each of the ten BMAbs in a competition-based ELISA. Five of the neutralizing BMAbs (C48, C65, C74, C83 and MH6) were shown to be directed against epitopes on, or in close proximity to, that previously defined as neutralizing antigenic site 2. Another neutralizing BMAb, MH5, bound to an epitope on, or in close proximity to antigenic site 3. Two neutralizing BMAbs (C2 and C96) simultaneously abrogated the binding of mouse antibodies to sites 2 and 4, contesting the autonomous nature of these two regions. None of the BMAbs were shown to be directed towards the immunodominant antigenic site 1. Sequence analyses of neutralization escape mutants supported the competition ELISA results, and included changes at site 2 (VP2 aa C78Y), site 3 (VP1 N46S) and site 4 (VP3 E58K). Additionally, a substitution at a previously unrecorded location (VP2 aa T1881) prevented the binding of site 2 (C48) and sites 2 and 4 (C2) directed BMAbs. Although the bovine and murine anti-FMDV repertoires may not be identical these results support the recognition of similar antigenic features. This is the first report characterizing anti-FMDV MAbs produced from a natural target host, the cow.
8种具有中和作用和2种不具有中和作用的抗口蹄疫病毒(FMDV)牛IgG1和IgG2单克隆抗体(BMAb)识别构象依赖性表位。大多数已证明具有中和病毒作用的抗体,无论其亚型如何,都能被动保护小鼠免受病毒攻击。在基于竞争的ELISA中,用10种BMAb分别检测了表征良好的抗FMDV小鼠单克隆抗体,这些小鼠单克隆抗体代表O1血清型病毒上的5个独立中和表位。其中5种具有中和作用的BMAb(C48、C65、C74、C83和MH6)被证明针对的是先前定义为中和抗原位点2上或其附近的表位。另一种具有中和作用的BMAb,MH5,与抗原位点3上或其附近的表位结合。2种具有中和作用的BMAb(C2和C96)同时消除了小鼠抗体与位点2和位点4的结合,对这两个区域的自主性提出了质疑。没有一种BMAb被证明针对免疫显性抗原位点1。中和逃逸突变体的序列分析支持了竞争ELISA结果,包括位点2(VP2第78位氨基酸由C变为Y)、位点3(VP1第46位氨基酸由N变为S)和位点4(VP3第58位氨基酸由E变为K)的变化。此外,在一个先前未记录的位置(VP2第1881位氨基酸)发生的取代阻止了针对位点2(C48)以及针对位点2和位点4(C2)的BMAb的结合。尽管牛和小鼠的抗FMDV抗体库可能不完全相同,但这些结果支持了对相似抗原特征的识别。这是第一篇描述从天然靶宿主牛产生的抗FMDV单克隆抗体的报告。