Yamamoto M, Black J B, Stewart J A, Lopez C, Pellett P E
Division of Viral and Rickettsial Diseases, Centers for Disease Control, Atlanta, Georgia 30333.
J Clin Microbiol. 1990 Sep;28(9):1957-62. doi: 10.1128/jcm.28.9.1957-1962.1990.
Enveloped whole virions and nucleocapsids of human herpesvirus 6 (HHV-6) strain Z29 were purified from supernatant fluids of infected human cord blood lymphocytes by filtration through polyvinylpyrrolidone-treated filters, banding on a Nycondenz step gradient, and centrifugation through two successive continuous sucrose gradients. More than 20 proteins ranging in molecular weight from less than 30,000 to more than 200,000 were identified in preparations of purified whole virions labeled with [35S]methionine and [35S]cysteine. Immunogenic virion proteins of HHV-6 were identified in immunoblot assays with human immune sera, immune sera generated from mice immunized with purified whole virions or purified nucleocapsids, and a monoclonal antibody generated from a mouse immunized with purified nucleocapsids. The sera and the monoclonal antibody reacted strongly with a 101-kilodalton protein in the immunoblots, suggesting that the protein is a component of the nucleocapsid. Human sera lacking HHV-6-specific antibodies and seropositive for one or more of the other human herpesviruses failed to react with this protein, indicating that it is a specific serologic marker for HHV-6 infection.
通过经聚乙烯吡咯烷酮处理的滤器过滤、在Nycondenz阶梯梯度上进行区带分离以及通过两个连续的连续蔗糖梯度离心,从感染的人脐血淋巴细胞的上清液中纯化出人类疱疹病毒6型(HHV - 6)Z29株的包膜完整病毒粒子和核衣壳。在用[35S]甲硫氨酸和[35S]半胱氨酸标记的纯化完整病毒粒子制剂中鉴定出20多种分子量从小于30,000到大于200,000的蛋白质。在用人类免疫血清、用纯化完整病毒粒子或纯化核衣壳免疫的小鼠产生的免疫血清以及用纯化核衣壳免疫的小鼠产生的单克隆抗体进行的免疫印迹分析中鉴定出HHV - 6的免疫原性病毒粒子蛋白。这些血清和单克隆抗体在免疫印迹中与一种101千道尔顿的蛋白质强烈反应,表明该蛋白质是核衣壳的一个组成部分。缺乏HHV - 6特异性抗体且对一种或多种其他人类疱疹病毒呈血清阳性的人血清未能与该蛋白质反应,表明它是HHV - 6感染的一种特异性血清学标志物。