Cornaglia E M, Elazhary Y M, Brodeur B R, Talbot B G
Virology Section, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, Quebec, Canada.
J Virol. 1992 Oct;66(10):5763-9. doi: 10.1128/JVI.66.10.5763-5769.1992.
This study describes, for the first time, the production and use of an "internal-image" anti-idiotypic monoclonal antibody (MAb) to elicit a rotavirus-specific antibody response. An immunoglobulin G2a MAb, designated RQ31 (MAb1), specific for the outer capsid protein VP4 of bovine Q17 rotavirus and capable of neutralizing viral infection in vitro was used to generate an anti-idiotypic MAb (MAb2). This MAb2, designated RQA2, was selected by enzyme-linked immunosorbent assay (ELISA) using F(ab')2 fragments of RQ31. RQA2 (MAb2) inhibited the binding of RQ31 (MAb1) to the virus but had no effect on the binding of other rotavirus-specific MAbs. The MAb2 also inhibited virus neutralization mediated by MAb1 in a dose-dependent fashion. Naive guinea pigs immunized with the MAb2 produced anti-anti-idiotypic antibodies (Ab3) that reacted with bovine Q17 rotavirus in an ELISA and neutralized rotavirus infection in vitro. The Ab3 response was characterized as MAb1-like because the Ab3 recognizes only the Q17 and neonatal calf diarrhea virus rotavirus strains in ELISA, as did RQ31 (MAb1). The Ab3 response also possessed two other characteristics of RQ31: the abilities to bind the 1.36 (double-capsid) but not the 1.38 (single-capsid) purified rotavirus fraction in ELISA and to immunoprecipitate the VP4 rotavirus protein.
本研究首次描述了一种“内影像”抗独特型单克隆抗体（MAb）的产生及其用于引发轮状病毒特异性抗体反应的情况。一种免疫球蛋白G2a单克隆抗体，命名为RQ31（MAb1），对牛Q17轮状病毒的外衣壳蛋白VP4具有特异性，且能够在体外中和病毒感染，用其产生了一种抗独特型单克隆抗体（MAb2）。这种MAb2命名为RQA2，是通过酶联免疫吸附测定（ELISA）使用RQ31的F(ab')2片段筛选出来的。RQA2（MAb2）可抑制RQ31（MAb1）与病毒的结合，但对其他轮状病毒特异性单克隆抗体的结合没有影响。MAb2还以剂量依赖性方式抑制MAb1介导的病毒中和作用。用MAb2免疫的未免疫豚鼠产生了抗抗独特型抗体（Ab3），这些抗体在ELISA中与牛Q17轮状病毒发生反应，并在体外中和轮状病毒感染。Ab3反应被表征为类似MAb1，因为在ELISA中Ab3仅识别Q17和新生小牛腹泻病毒轮状病毒株，RQ31（MAb1）也是如此。Ab3反应还具有RQ31的另外两个特征：在ELISA中能够结合1.36（双链体）而非1.38（单链体）纯化轮状病毒组分，以及能够免疫沉淀轮状病毒VP4蛋白。
