IDENTIFICATION OF A 5TH NEUTRALIZABLE SITE ON TYPE-O FOOT-AND-MOUTH-DISEASE VIRUS FOLLOWING CHARACTERIZATION OF SINGLE AND QUINTUPLE MONOCLONAL-ANTIBODY ESCAPE MUTANTS
IDENTIFICATION OF A 5TH NEUTRALIZABLE SITE ON TYPE-O FOOT-AND-MOUTH-DISEASE VIRUS FOLLOWING CHARACTERIZATION OF SINGLE AND QUINTUPLE MONOCLONAL-ANTIBODY ESCAPE MUTANTS
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DOI：
10.1099/0022-1317-74-8-1547
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发表时间：
1993-08-01
影响因子：
3.8
通讯作者：
SAMUEL, AR
中科院分区：
文献类型：
--
作者：
CROWTHER, JR;FARIAS, S;SAMUEL, AR
发现一种抗口蹄疫病毒O1Caseros型单克隆抗体（C3）能中和口蹄疫病毒O1Kaufbeuren型四价单克隆抗体逃逸突变体（G67）。该突变体在序列水平上具有明显的变化，影响了四个不重叠的可中和位点。用C3单克隆抗体制备G67病毒的五重逃逸突变体和亲本O1Kaufbeuren病毒的单逃突变体。多克隆接种和感染后的牛血清以及多克隆小鼠和豚鼠血清，中和了四价突变体，不再中和五价突变体，这表明已经确定了第五个位点，改变第五个位点消除了所有中和作用。使用血清学技术对该位点进行了表征，发现该位点具有构象依赖性，胰蛋白酶敏感性，并且独立于先前由单克隆抗体表征的位点。对亲本突变体、C3突变体和5倍突变体的氨基酸测序结果显示，VP1结构蛋白的第149个氨基酸发生了从谷氨酰胺到组氨酸的单一变化。（1D）表征C3突变。第五个位点可能是由于VP1环区与其他表面氨基酸相互作用而形成的构象表位。
A monoclonal antibody (C3) produced against foot-and-mouth disease virus type O1Caseros was found to neutralize quadrivalent monoclonal antibody escape mutant (G67) of foot-and-mouth disease virus type O1Kaufbeuren. This mutant had been characterized at the sequence level as having distinct changes affecting four non-overlapping neutralizable sites. The C3 monoclonal antibody was used to prepare a quintuple escape mutant from the G67 and a single escape mutant from the parental O1Kaufbeuren viruses. Polyclonal postvaccinated and infected cattle sera as well as polyclonal mouse and guinea-pig sera, which neutralized the quadrivalent mutant, no longer neutralized the quintuple mutant, indicating that a fifth site had been identified and that changing the fifth site eliminated all neutralization. The site was characterized using serological techniques and found to be conformationally dependent, trypsin-sensitive and independent of sites previously characterized by monoclonal antibodies. Amino acid sequencing comparing parental, single C3 and quintuple mutants showed that a single change from a glutamine to a histidine, at amino acid 149 in the structural protein VP1. (1D) characterized the C3 mutation. The fifth site probably represents a conformational epitope which is formed due to the interaction of the VP1 loop region with other surface amino acids.