Neutralizing monoclonal antibody to the E1 glycoprotein epitope of rubella virus mediates virus arrest in VERO cells
Neutralizing monoclonal antibody to the E1 glycoprotein epitope of rubella virus mediates virus arrest in VERO cells
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DOI：
10.1089/vim.2000.13.83
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发表时间：
2000-01-01
期刊：
影响因子：
2.2
通讯作者：
Zapata, M
中科院分区：
文献类型：
--
作者：
Corboba, P;Grutadauria, S;Zapata, M
抗体介导的病毒中和作用的最著名的作用机制是阻止病毒进入宿主细胞，如通过中和测定所确定的。抗体也可以抑制风疹病毒（RV）从感染的宿主细胞中退出;在这种情况下，抗体与其结构域的相互作用必须发生在质膜上，因为抗体不能进入细胞。在本研究中，我们能够暂时阻止退出的病毒粒子从RV感染的细胞的单克隆抗体（mAb）H3的结合到他们的表面。该目标通过三个步骤实现：首先，我们确定了病毒复制周期的持续时间;然后，我们建立了RV感染的VERO细胞的细胞质中由我们的mAb定义的结构域的存在的动力学;最后，我们评估了在存在或不存在mAb或阳性和阴性小鼠血清的情况下，病毒颗粒释放到感染的VERO细胞的上清液中。RV特异性小鼠血清和与RV-E1糖蛋白的氨基酸序列208-239结合的mAb H3能够将病毒体从感染的培养物中释放延迟24小时，这表明mAb H3与其表位的反应可以阻止病毒体组装和/或释放所需的任何变化。总之，mAb识别的中和结构域诱导的抗体可以通过多种作用机制阻断病毒复制，例如阻断病毒进入细胞和延迟病毒释放。所有这些机制都密切参与了关键的病毒-宿主细胞相互作用，使感染的自我限制。
The best-known mechanism of action of antibody-mediated virus neutralization is to impede the entrance of viruses to host cells, as determined by neutralization assays. Antibodies may also inhibit the exit of rubella virus (RV) from infected host cells; in this case, the interaction of the antibodies with their domains must occur on the plasma membrane, because antibodies cannot enter the cells. In the present study, we were able to block temporally the exit of virions from RV-infected cells by the binding of monoclonal antibody (mAb) H3 to their surface. The objective was accomplished in three steps: first, we determined the duration of the viral replication cycle; then we established the kinetics of the presence of the domains defined by our mAbs in the cytoplasm of RV-infected VERO cells; and, finally, we assessed the release of viral particles to the supernatant of infected VERO cells in the presence or absence of mAbs or positive and negative mice sera. RV-specific mice sera and mAb H3, which binds to the amino acid sequence 208-239 of the RV-E1 glycoprotein, were able to delay for 24 hours the release of virions from infected cultures, suggesting that the reaction of mAb H3 with its epitope may arrest any change necessary for the assembly and/or release of virions. In conclusion, the neutralizing domain recognized by mAb induces antibodies that can block the viral replication by several mechanisms of action, such as the obstruction of virus entry into cells and the delay of viral release. All of these mechanisms are intimately involved in the critical virus-host cell interactions that allow self-limitation of the infection.