Generation and characterization of mouse monoclonal antibodies against the VP4 protein of group A human rotaviruses
Generation and characterization of mouse monoclonal antibodies against the VP4 protein of group A human rotaviruses
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DOI：
10.1016/j.antiviral.2022.105407
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发表时间：
2022-09-26
影响因子：
7.6
通讯作者：
Xia，Ningshao
中科院分区：
文献类型：
--
作者：
Zeng，Yuanjun;Song，Feibo;Xia，Ningshao
人类轮状病毒（RVs）是全世界婴幼儿严重腹泻的主要原因。在结构蛋白中，轮状病毒VP4作为一种刺突蛋白，在病毒的附着和渗透过程中都起着关键作用。目前，针对VP4的单克隆抗体（mab）研究有限。在这项研究中，用截断的VP4*免疫小鼠产生小鼠单克隆抗体。共制备和鉴定了50个单抗。24个单抗具有基因型特异性，20个单抗识别P[8]、P[4]和P[6]病毒共有的VP4表位。50个单抗中有35个是中和性单抗，其中9个单抗能中和所有3种p基因型rv， 10个中和性单抗表现出构象敏感性。10个单抗识别显性中和表位，包括广泛中和的单抗9C4识别构象表位。进一步研究表明，S376和S464是9C4结合的关键氨基酸，但9C4的确切结合位点仍未完全确定。总的来说，这组单克隆抗体已经证明了作为免疫诊断和研究试剂的实用性，并且可能作为探索基于VP4*蛋白的RV亚单位疫苗的中和机制和质量控制的重要工具。进一步评价交叉中和单克隆抗体不仅可以提高对RV疫苗异型保护作用的认识，而且有助于开发具有广泛保护作用的RV疫苗。
Human rotaviruses (RVs) are the leading cause of severe diarrhea in infants and young children worldwide. Among the structural proteins, as a spike protein, rotavirus VP4 plays a key role in both viral attachment and penetration. Currently, studies on monoclonal antibodies (mAbs) against VP4 are limited. In this study, mice were immunized with truncated VP4* to produce murine mAbs. In total, 50 mAbs were produced and characterized. Twenty-four mAbs were genotype-specific and 20 mAbs recognized the common VP4 epitopes shared by P[8], P[4], and P[6] viruses. Thirty-five of the 50 mAbs were neutralizing mAbs, among which nine mAbs could neutralize all three P-genotype RVs, and 10 neutralizing mAbs exhibited conformational sensitivity. Ten mAbs recognized dominant neutralizing epitopes, including the broadly neutralizing mAb 9C4 recognized conformational epitope. Further investigation shows that S376 and S464 are key amino acids for 9C4 binding, however, the exact binding sites of 9C4 remain to be fully defined. Overall, this panel of mAbs has demonstrated utility as immunodiagnostic and research reagents, and could potentially serve as crucial tools for exploring the neutralizing mechanisms and quality control of VP4* protein-based RV subunit vaccines. Further evaluation of cross-neutralizing mAbs could not only improve the understanding of the heterotypic protection conferred by RV vaccines, but also facilitate the development of broadly protective RV vaccines.