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

1个血清型特异性和3个血清型交叉反应中和 用单抗(N-mAbs)研究中和作用 抗原表位参与了抗原位点的形成。单人，双人， 或三重中和抗性突变体用N- 单抗在体外按顺序排列。核苷酸和推导的氨基酸(AA) 这些突变体的VP7上的替换表明：(I)在 除了可变区VR-5(AA 87-100)、VR-7(AA 141-150)、VR-8 (AA 208-224)和VR-9(AA 235-242)，AA 290和291恒定 VP7区参与中和，以及(Ii)VR-5、VR-7、 VR-8、VR-9以及AA 290和291在功能上彼此相关。 为了进一步分析VP7上的中和点并研究 所用N-单抗之间可能的协同或拮抗作用 在这项研究中，在三种病毒存在的情况下产生了抗原变体 N-单抗。猪瘟病毒VP7编码基因的核苷酸序列分析 突变体不显示单抗之间的协同或拮抗作用 用于筛选中和逃逸突变体。

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.