Department of Environmental Health, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
J Gen Virol. 2014 Jan;95(Pt 1):171-178. doi: 10.1099/vir.0.055608-0. Epub 2013 Oct 4.
Enteric human adenoviruses (HAdVs; serotypes 40 and 41) have been identified as an emerging cause of drinking water contamination. Due to their fastidious characteristics, HAdVs are difficult to cultivate and therefore not detected easily by standard mammalian cell cultivation methods. Here we found that human embryonic kidney 293 cells, transformed transiently with Ras, enhanced HAdV replication by more than threefold. We also constructed a stable cell line overexpressing the Ras protein, 293-Ras, in which the replication of three HAdV strains of serotypes 40 and 41 was increased markedly. However, only HAdV replication was enhanced; infection of 293 and 293-Ras cells with human rhinivorus-6 showed no significant differences in replication rate. Infected 293-Ras cells exhibited an increased level and phosphorylation of extracellular regulated kinase (ERK). In addition, the Ras-mediated increase in HAdV replication was impaired by the mitogen-activated protein kinase/ERK kinase (MEK1) inhibitor U0126, suggesting direct involvement of the MEK1/ERK pathway in enhanced HAdV replication. Based on these results, we suggest that the 293-Ras cell line be used for the efficient detection and cultivation of HAdV strains in both clinical and environmental specimens.
肠道人类腺病毒（HAdV；血清型 40 和 41）已被确定为饮用水污染的一个新出现的原因。由于其苛刻的特性，HAdV 难以培养，因此不易通过标准的哺乳动物细胞培养方法检测到。在这里，我们发现瞬时转化 Ras 的人胚肾 293 细胞使 HAdV 复制增加了三倍以上。我们还构建了一个稳定表达 Ras 蛋白的细胞系 293-Ras，其中三种血清型 40 和 41 的 HAdV 株的复制明显增加。然而，只有 HAdV 复制得到增强；感染 293 和 293-Ras 细胞的人类鼻病毒-6 的复制率没有明显差异。感染的 293-Ras 细胞表现出细胞外调节激酶（ERK）水平和磷酸化的增加。此外，Ras 介导的 HAdV 复制增加被丝裂原活化蛋白激酶/ERK 激酶（MEK1）抑制剂 U0126 所抑制，这表明 MEK1/ERK 途径直接参与了增强的 HAdV 复制。基于这些结果，我们建议使用 293-Ras 细胞系来有效检测和培养临床和环境标本中的 HAdV 株。
