Institute of Health and Environment, Department of Environmental Health, School of Public Health, Seoul National University, 28 Yeongeon-dong, Chongro-gu, Seoul, South Korea 110-799.
Appl Environ Microbiol. 2010 Apr;76(8):2509-16. doi: 10.1128/AEM.02224-09. Epub 2010 Feb 5.
Human enteric adenoviruses (HAdVs; serotypes 40 and 41) are important waterborne and food-borne pathogens. However, HAdVs are fastidious, are difficult to cultivate, and do not produce a clear cytopathic effect during cell culture within a reasonable time. Thus, we examined whether the viral transactivator proteins cytomegalovirus (CMV) IE1 and hepatitis B virus (HBV) X promoted the multiplication of HAdVs. Additionally, we constructed a new 293 cell line expressing CMV IE1 protein for cultivation assays. We analyzed the nucleic acid sequences of the promoter regions of both E1A and hexon genes, which are considered to be the most important regions for HAdV replication. Expression of either HBV X or CMV IE1 protein significantly increased the promoter activities of E1A and hexon genes of HAdVs by as much as 14-fold during cell cultivation. The promotion of HAdV expression was confirmed by increased levels of both adenoviral DNA and mRNA expression. Finally, the newly developed 293 cell line expressing CMV IE1 protein showed an increase in viral DNA ranging from 574% to 619% compared with the conventional 293 cell line. These results suggest that the newly constructed cell line could be useful for efficient cultivation and research of fastidious HAdVs.
人肠道腺病毒（HAdV；血清型 40 和 41）是重要的水源性和食源性病原体。然而，HAdV 是苛刻的，难以培养，并且在合理的时间内不会在细胞培养中产生明显的细胞病变效应。因此，我们研究了巨细胞病毒（CMV）IE1 和乙型肝炎病毒（HBV）X 等病毒转录激活蛋白是否能促进 HAdV 的增殖。此外，我们构建了一种新的表达 CMV IE1 蛋白的 293 细胞系，用于培养实验。我们分析了 E1A 和 hexon 基因启动子区域的核酸序列，这些区域被认为是 HAdV 复制的最重要区域。在细胞培养过程中，HBV X 或 CMV IE1 蛋白的表达分别使 HAdV 的 E1A 和 hexon 基因的启动子活性增加了 14 倍。通过增加腺病毒 DNA 和 mRNA 表达水平，证实了 HAdV 表达的促进。最后，与传统的 293 细胞系相比，新构建的表达 CMV IE1 蛋白的 293 细胞系中的病毒 DNA 水平增加了 574%至 619%。这些结果表明，新构建的细胞系可用于高效培养和研究苛刻的 HAdV。
