ANALYSIS OF NEUTRALIZING EPITOPES ON ROTAVIRUS OUTER CAPSID PROTEINS (VP4, VP7)

一型特异三型交叉反应中和 单克隆抗体（N-mAbs）用于研究中和 参与抗原位点形成的表位。 单人，双人， 或三重中和抗性突变体通过使用N- mAb体外顺序。 核苷酸和推导的氨基酸（aa） 在此类突变体的VP 7上发现的取代显示：（i）在 除了可变区VR-5（aa 87-100）、VR-7（aa 141-150）、VR-8（aa 141-150）之外， (aa 208-224），和VR-9（aa 235-242），aa 290和291在常数 VP 7的区域参与中和，和（ii）VR-5，VR-7， VR-8、VR-9和aa 290和291在功能上彼此相关。 为了进一步分析VP 7上的中和位点并研究 所用N-mAb之间可能的协同或拮抗作用 在这项研究中，抗原变体是在三种 N-mAb。 日本血吸虫VP 7编码基因的核苷酸序列分析 突变体显示mAb之间没有协同或拮抗作用 用于选择中和逃逸突变体。

One serotype-specific and three serotype-cross-reactive neutralizing monoclonal antibodies (N-mAbs) were utilized to study the neutralization epitopes involved in the formation of antigenic sites. Single, double, or triple neutralization-resistant mutants were selected by using the N- mAbs sequentially in vitro. Nucleotide and deduced amino acid (aa) substitutions found on the VP7 of such mutants showed that: (i) in addition to variable regions VR-5 (aa 87-100), VR-7 (aa 141-150), VR-8 (aa 208-224), and VR-9 (aa 235-242), aa 290 and 291 in the constant region of the VP7 are involved in neutralization, and (ii) VR-5, VR-7, VR-8, VR-9, and aa 290 and 291 are functionally related to one another. In order to further analyze neutralization sites on the VP7 and to study possible synergistic or antagonistic effects among the N-mAbs employed in this study, antigenic variants were generated in the presence of three N-mAbs. Nucleotide sequence analysis of the VP7-encoding gene of the mutants revealed no synergistic or antagonistic effects among the mAbs used in the selection of neutralization escape mutants.