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

轮状病毒外衣壳蛋白（VP4、VP7）中和表位的分析

• 批准号: 5200542
• 负责人: Y HOSHINO
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5200542
• 关键词: Rotavirus; antiviral antibody; capsid; epitope mapping; gene mutation; microorganism immunology; monoclonal antibody; neutralizing antibody; nucleic acid sequence; protein sequence; virus antigen; virus protein

One serotype-specific and three serotype-cross-reactive anti-VP7 neutralizing monoclonal antibodies (N-mAbs) were utilized to study the neutralization epitopes involved in the formation of antigenic sites on porcine rotavirus Gottfried strain (serotype 4). Single, double, or triple neutralization-resistant mutants were selected in the presence of these N-mAbs in vitro. Sequence analysis of the gene encoding VP7 of such mutants revealed that: (i) the amino acid (aa) that is substituted at a specific position on the mutant VP7 selected by a single N-mAb can vary, resulting in variants which exhibit antigenic differences; (ii) 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 (iii) 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 failed to demonstrate synergistic or antagonistic effects among the mAbs used in the selection of neutralization escape mutants.

1种亚型特异性和3种亚型交叉反应性抗VP 7 中和性单克隆抗体（N-mAbs）用于研究 中和表位参与抗原位点的形成 猪轮状病毒Gottfried株（血清型4）。单人间，双人间，还是 三重中和抗性突变体是在以下存在下选择的： 这些N-mAb在体外。编码这种病毒的VP 7基因的序列分析 突变体揭示：（i）在A处被取代的氨基酸（aa）， 由单个N-mAb选择的突变体VP 7上的特定位置可以变化， 导致表现出抗原差异的变体;（ii）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），恒定区中的aa 290和291 的VP 7参与中和，和（iii）VR-5，VR-7，VR-8， VR-9和aa 290和291在功能上彼此相关。在 为了进一步分析VP 7上的中和位点， 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 突变体未能表现出协同或拮抗作用， 用于选择中和逃逸突变体的mAb。