RRC ID 68628
著者 Kawasaki H, Kosugi I, Arai Y, Iwashita T, Tsutsui Y.
タイトル Mouse embryonic stem cells inhibit murine cytomegalovirus infection through a multi-step process.
ジャーナル PLoS One
Abstract In humans, cytomegalovirus (CMV) is the most significant infectious cause of intrauterine infections that cause congenital anomalies of the central nervous system. Currently, it is not known how this process is affected by the timing of infection and the susceptibility of early-gestational-period cells. Embryonic stem (ES) cells are more resistant to CMV than most other cell types, although the mechanism responsible for this resistance is not well understood. Using a plaque assay and evaluation of immediate-early 1 mRNA and protein expression, we found that mouse ES cells were resistant to murine CMV (MCMV) at the point of transcription. In ES cells infected with MCMV, treatment with forskolin and trichostatin A did not confer full permissiveness to MCMV. In ES cultures infected with elongation factor-1α (EF-1α) promoter-green fluorescent protein (GFP) recombinant MCMV at a multiplicity of infection of 10, less than 5% of cells were GFP-positive, despite the fact that ES cells have relatively high EF-1α promoter activity. Quantitative PCR analysis of the MCMV genome showed that ES cells allow approximately 20-fold less MCMV DNA to enter the nucleus than mouse embryonic fibroblasts (MEFs) do, and that this inhibition occurs in a multi-step manner. In situ hybridization revealed that ES cell nuclei have significantly less MCMV DNA than MEF nuclei. This appears to be facilitated by the fact that ES cells express less heparan sulfate, β1 integrin, and vimentin, and have fewer nuclear pores, than MEF. This may reduce the ability of MCMV to attach to and enter through the cellular membrane, translocate to the nucleus, and cross the nuclear membrane in pluripotent stem cells (ES/induced pluripotent stem cells). The results presented here provide perspective on the relationship between CMV susceptibility and cell differentiation.
巻・号 6(3)
ページ e17492
公開日 2011-3-2
DOI 10.1371/journal.pone.0017492
PMID 21407806
PMC PMC3047572
MeSH Animals Cell Differentiation / drug effects Cell Line Centrifugation Colforsin / pharmacology Cytomegalovirus Infections / metabolism* Cytomegalovirus Infections / virology* Embryo, Mammalian / cytology Embryonic Stem Cells / cytology Embryonic Stem Cells / drug effects Embryonic Stem Cells / metabolism Embryonic Stem Cells / virology* Fibroblasts / drug effects Fibroblasts / metabolism Genes, Immediate-Early Genome, Viral / genetics Hydroxamic Acids / pharmacology In Situ Hybridization Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / virology Mice Mice, Inbred C57BL Muromegalovirus / drug effects Muromegalovirus / genetics Muromegalovirus / pathogenicity Muromegalovirus / physiology* Peptide Elongation Factor 1 / metabolism Promoter Regions, Genetic / genetics Recombination, Genetic / genetics Transfection
IF 2.74
リソース情報
ヒト・動物細胞 iPS-MEF-Ng-20D-17(APS0001)