Production of a monoclonal antibody against the Snow Mountain agent of gastroenteritis by in vitro immunization of murine spleen cells.
Production of a monoclonal antibody against the Snow Mountain agent of gastroenteritis by in vitro immunization of murine spleen cells.
复制标题
通过小鼠脾细胞体外免疫制备针对雪山胃肠炎病原体的单克隆抗体。
DOI：
10.1073/pnas.85.10.3613
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
Madore,HP
中科院分区：
文献类型：
--
作者：
Treanor,J;Dolin,R;Madore,HP
雪山病毒（SMA）是一种27- 32纳米的不可培养病毒，可引起人类急性胃肠炎。SMA在形态学上与诺瓦克菌相似，但在免疫学上与诺瓦克菌不同。SMA已从实验感染志愿者的粪便中部分纯化，含有Mr 62,000的单一结构蛋白以及一种或多种非病毒相关的可溶性蛋白。这种重要的人类病原体和其他诺瓦克样病毒的进一步表征由于缺乏研究它们的试剂而受到阻碍。为了进一步表征SMA，我们开发了一种使用体外免疫的SMA单克隆抗体，这种技术允许使用少量抗原进行免疫。单克隆抗体SM-4对SMA具有特异性，与诺沃克或夏威夷制剂不发生反应。此外，SM-4与纯化的病毒粒子反应，但不与可溶性蛋白反应。SM-4还阻断了标记的感染后人IgG与纯化病毒粒子结合的能力。最后，SM-4和人感染后血清特异性识别Mr 62000病毒相关蛋白。因此，SM-4针对存在于SMA结构蛋白上的表位，该表位不为诺沃克或夏威夷试剂所共享，也不存在于可溶性蛋白上。抗SMA单克隆抗体的可用性将有助于进一步纯化和表征该药物。这些研究中使用的技术为生产针对这组病毒的额外单克隆抗体提供了一种方法，也应该对其他隐匿病毒制剂的研究有用。
The Snow Mountain agent (SMA) is a 27- to 32-nm noncultivatable virus that causes acute gastroenteritis in humans. SMA is morphologically similar to but immunologically distinct from the Norwalk agent. SMA has been partially purified from the stool of experimentally infected volunteers and contains a single structural protein of Mr 62,000 as well as one or more non-virion-associated soluble proteins. Further characterization of this important human pathogen and other Norwalk-like viruses has been hindered by the lack of reagents with which to study them. To further characterize SMA, we developed a monoclonal antibody to SMA using in vitro immunization--a technique that permitted use of small quantities of antigen for immunization. The monoclonal antibody, SM-4, was specific for SMA and did not react with the Norwalk or Hawaii agents. In addition, SM-4 reacted with purified virion but not with the soluble protein. SM-4 also blocked the ability of labeled postinfection human IgG to bind to purified virion. Finally, both SM-4 and human postinfection sera specifically recognized the Mr 62,000 virion-associated protein. Thus, SM-4 is directed against an epitope present on the SMA structural protein that is not shared by the Norwalk or Hawaii agents and that is not present on the soluble protein. The availability of a monoclonal antibody against SMA should facilitate further purification and characterization of this agent. The techniques utilized in these studies provide a method for the production of additional monoclonal antibodies to this group of viruses and also should be useful for the study of other occult viral agents.