In-vitro cultivation of human cytomegalovirus in thyroid epithelial cells

Knowles, Wendy A.

doi:10.1007/bf01318006

Cited by 27 publications

(14 citation statements)

References 12 publications

(5 reference statements)

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“…The virus has a large linear DNA genome of 240 kbp which is organized into long and short unique components that invert to give four sequence isomers (LaFemina & Hayward, 1980;Fleckenstein et al, 1982). I-ICMV displays a strict host cell type and species specificity (Rowe et al, 1956;Smith, 1956), although several reports have shown that HCMV replicates to a limited degree in human epithelial ceils (MichelsonFiske et al, 1975;Knowles, 1976;Vonka et al, 1976). Following the infection of susceptible ceils with HCMV, expression of the viral genome proceeds in three main phases: immediate early (IE), early and late (Stinski et al, 1980;DeMarchi, 1981;McDonough & Spector, 1983;Griffiths & Grundy, 1987).…”

Section: Introductionmentioning

confidence: 99%

Differential Expression of the Major Immediate Early Gene of Human Cytomegalovirus

Tsutsui

Nogami-Satake²

1990

Journal of General Virology

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We prepared a murine monoclonal antibody reactive to a human cytomegalovirus (HCMV)-induced nuclear protein with an Mr of 68000. Expression of the 68K protein was compared with the major immediate early (IE) 72K protein in various cell types after infection with HCMV or microinjection of plasmid DNA containing the major IE gene. The 68K nuclear protein was detected 2 to 3 h after appearance of the 72K protein in human embryonal lung (HEL) cells infected with HCMV. The 68K protein was distributed throughout the cytoplasm in the late phase of infection, while the 72K protein remained chiefly in the nucleus. The 68K protein was barely detected in the cells under IE conditions by immunoprecipitation, but, together with the 72K protein, it was expressed after microinjection of cloned DNA, containing only the major IE region (region 1), into the nuclei of HEL cells. The 72K protein was expressed in nuclei 2 h after microinjection, whereas the 68K protein was detected 4 to 5 h after the injection. The 68K protein was expressed after microinjection in non-permissive rodent fibroblasts or non-permissive transformed human cells in which these proteins were not expressed after viral infection.Immunoprecipitations after chase-labelling from IE conditions or after partial digestions suggested that the 68K protein is neither a degradation nor a modification product of the major IE 72K protein.

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Section: Introductionmentioning

confidence: 99%

Differential Expression of the Major Immediate Early Gene of Human Cytomegalovirus

Tsutsui

Nogami-Satake²

1990

Journal of General Virology

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“…Other human cell types tested such as epithelial cells are, at best, only moderately permissive (Knowles, 1976;Vonka et aL, 1976). However, efficient growth even in fibroblast cultures apparently requires adaptation by serial passaging in cell culture.…”

Section: Introductionmentioning

confidence: 99%

Differences in Cell Type-specific Blocks to Immediate Early Gene Expression and DNA Replication of Human, Simian and Murine Cytomegalovirus

LaFemina

Hayward

1988

Journal of General Virology

100

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SUMMARYWe have previously described blocks to the viral lytic cycle at two different levels in cytomegalovirus (CMV)-infected non-permissive cells. BALB/c-3T3 cells express only the predominant immediate early (IE) nuclear phosphoproteins (IE68 or IE94) of human CMV (HCMV) or simian CMV (SCMV) and do not replicate the input viral genomes. However, in human teratocarcinoma stem cells and 293 cells, expression of the HCMV IE68 gene (but not the SCMV IE94 gene) is blocked at the transcriptional level. Here we report the results of an extensive comparison of the level of permissiveness for HCMV, SCMV and murine CMV (MCMV) in a variety of additional cell types of human, monkey and mouse origin. We also describe a subtle change in the tryptic peptide pattern of the IE68 polypeptide produced in BALB/c-3T3 cells compared to permissive human foreskin fibroblasts. Neither the IE68 nor IE94 proteins could be detected by biochemical labelling procedures in infected mouse Ltkor F9 teratocarcinoma stem cells, although IE94 was synthesized after retinoic acidinduced differentiation of the F9 cells. Synthesis of [35S]methionine-labelled IE94 protein, but not that of HCMV IE68, was detected in infected Vero cells and in human peripheral blood leukocyte cultures. The failure to synthesize detectable IE68 protein in infected Vero cells appeared to be unrelated to a lack of entry of viral DNA and to a lack of appropriate transcription factors. Indeed, immunofluorescence assays showed that the IE68 antigen was expressed efficiently in DNA-transfected Vero cells and in a small fraction of infected Vero cells. Overall, two clear host range trends emerged. First, whilst all three viruses showed a tendency for repression of IE expression in transformed cell lines, the effect was severe for HCMV and only minimal for SCMV. Secondly, progression of infection to the viral DNA synthesis level in non-transformed fibroblast cell types occurred in a much wider range of host species cell types for SCMV and MCMV than for HCMV.

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“…Despite the frequent and extensive infection of epithelial cell populations by HCMV in vivo, only a few cultured epithelial cells have been reported to be susceptible to HCMV infection, including primary lung epithelial cells (30), thyroid epithelial cells (26), HCMC and Caco-2 epithelial cells derived from colon (24,40), cervical epithelial cells (41), and BAMB cells derived from amniotic fluid cells (42). However, most analyses of HCMV epithelial cell tropism have been performed using laboratory-adapted strains, and potential variations among virus strains were not taken into consideration.…”

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confidence: 99%

Human Cytomegalovirus UL131 Open Reading Frame Is Required for Epithelial Cell Tropism

Wang

Shenk

2005

J Virol

328

367

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In-vitro cultivation of human cytomegalovirus in thyroid epithelial cells

Cited by 27 publications

References 12 publications

Differential Expression of the Major Immediate Early Gene of Human Cytomegalovirus

Differential Expression of the Major Immediate Early Gene of Human Cytomegalovirus

Differences in Cell Type-specific Blocks to Immediate Early Gene Expression and DNA Replication of Human, Simian and Murine Cytomegalovirus

Human Cytomegalovirus UL131 Open Reading Frame Is Required for Epithelial Cell Tropism

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