Treanor J, Dolin R, Madore H P
Infectious Diseases Unit, University of Rochester School of Medicine and Dentistry, NY 14642.
Proc Natl Acad Sci U S A. 1988 May;85(10):3613-7. doi: 10.1073/pnas.85.10.3613.
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.
雪山因子(SMA)是一种27至32纳米、不可培养的病毒,可引起人类急性肠胃炎。SMA在形态上与诺沃克因子相似,但在免疫方面有所不同。SMA已从实验感染志愿者的粪便中部分纯化出来,含有一种分子量为62,000的单一结构蛋白以及一种或多种与非病毒体相关的可溶性蛋白。由于缺乏用于研究的试剂,对这种重要的人类病原体和其他诺沃克样病毒的进一步表征受到了阻碍。为了进一步表征SMA,我们使用体外免疫技术开发了一种针对SMA的单克隆抗体——该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,不与诺沃克或夏威夷因子发生反应。此外,SM - 4与纯化的病毒体发生反应,但不与可溶性蛋白发生反应。SM - 4还阻断了标记的感染后人类IgG与纯化病毒体结合的能力。最后,SM - 4和人类感染后血清都特异性识别分子量为62,000的病毒体相关蛋白。因此,SM - 4针对的是SMA结构蛋白上存在的一个表位,该表位既不为诺沃克或夏威夷因子所共有,也不存在于可溶性蛋白上。针对SMA的单克隆抗体的可用性应有助于对该病原体进行进一步的纯化和表征。这些研究中使用的技术为生产针对这组病毒的其他单克隆抗体提供了一种方法,也应有助于对其他隐匿性病毒病原体的研究。