DeMaula C D, Heidner H W, Rossitto P V, Pierce C M, MacLachlan N J
Department of Veterinary Pathology, School of Veterinary Medicine, University of California-Davis 95616.
Virology. 1993 Jul;195(1):292-6. doi: 10.1006/viro.1993.1377.
A panel of five neutralization-resistant escape mutant (EM) viruses was used to investigate the neutralization determinants of the U.S. prototype strain of bluetongue virus serotype 10 (BTV-10). The phenotypic properties of each EM virus were characterized by neutralization and immuneprecipitation assays with a panel of four monoclonal antibodies (MAbs). These MAbs were used to select the various EM viruses and together the MAbs define four distinct neutralizing epitopes on the prototype strain of BTV-10 (Heidner, H.W., Rositto, P.V., and MacLachlan, N.J., Virology 176, 658-661 (1990)). Sequencing of the L2 gene identified mutations responsible for the altered phenotypic properties exhibited by each EM virus. The L2 gene encodes BTV outer capsid protein VP2 which is responsible for virus neutralization. Four amino acids in three distinct regions of VP2 are critical to expression of the epitopes recognized by the MAb panel. Both amino acid 208 and 211 can affect the binding of MAb 039 and MAb 045, amino acid 327 affects binding of MAb 041, and amino acids 327 and 402 cooperatively interact to affect binding by MAb 034. The location of two of these critical regions on VP2 of BTV-10 is identical to two of those which affect neutralization of Australian BTV-1, despite the fact that these two viruses are antigenically distinct and have divergent L2 gene sequences (Gould, A.R., and Eaton, B.T., Virus Res. 17, 161-172 (1990)). The four individual neutralizing epitopes on VP2 of BTV-10 are interactive (Heidner, H.W., Rositto, P.V., and MacLachlan, N.J., Virology 176, 658-661 (1990)) and at least two are conformationally dependent.
使用一组五种抗中和逃逸突变体（EM）病毒来研究蓝舌病毒血清型10（BTV-10）美国原型株的中和决定因素。通过用一组四种单克隆抗体（MAb）进行中和和免疫沉淀试验，对每种EM病毒的表型特性进行了表征。这些单克隆抗体用于筛选各种EM病毒，并且这些单克隆抗体共同定义了BTV-10原型株上四个不同的中和表位（海德纳，H.W.，罗西托，P.V.，和麦克拉克伦，N.J.，《病毒学》176，658 - 661（1990））。对L2基因进行测序确定了导致每种EM病毒表现出改变的表型特性的突变。L2基因编码负责病毒中和的BTV外衣壳蛋白VP2。VP2三个不同区域中的四个氨基酸对于单克隆抗体组所识别表位的表达至关重要。氨基酸208和211都可影响单克隆抗体039和单克隆抗体045的结合，氨基酸327影响单克隆抗体041的结合，并且氨基酸327和402协同相互作用以影响单克隆抗体034的结合。尽管这两种病毒在抗原性上不同且L2基因序列不同，但BTV-10的VP2上这两个关键区域中的两个的位置与影响澳大利亚BTV-1中和的两个区域相同（古尔德，A.R.，和伊顿，B.T.，《病毒研究》17，161 - 172（1990））。BTV-10的VP2上的四个单独的中和表位是相互作用的（海德纳，H.W.，罗西托，P.V.，和麦克拉克伦，N.J.，《病毒学》176，658 - 661（1990）），并且至少两个是构象依赖性的。
