Human Cytomegalovirus Infections in vitro after Treatment with Arildone

Tyms, A. S.; Stevens, Robert; Mobberley, Margaret A.; Ryder, T. A.; Jeffries, D. J.

doi:10.1099/0022-1317-65-12-2129

Cited by 10 publications

(5 citation statements)

References 37 publications

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“…MRC-5 cells or the WF strain of skin/muscle embryonic fibroblasts (HEF cells) were grown and maintained as described previously (Tyms et al, 1984). The AD 169 strain of human cytomegalovirus was used in all experiments and cells were infected at a m.o.i, of 1 to l0 p.f.u./cell by adsorption for 2 h at 20 °C.…”

Section: Methodsmentioning

confidence: 99%

Fine Structure of Cells Infected with Human Cytomegalovirus After Treatment with 9-(1,3-Dihydroxy-2-propoxymethyl)guanine

Mobberley¹,

Ryder²,

Hart³

et al. 1987

Journal of General Virology

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SUMMARYInfection with human cytomegalovirus (CMV) is characterized by cytological changes which are readily visualized by electron microscopy using ultrathin sections of infected cells. Treatment of such cells with 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG), a potent inhibitor of CMV, is effective when initiated at early or late times after infection and the response to such treatment has been studied by fine structural analysis. Inhibition of viral DNA synthesis by DHPG treatment (50 laM) late in virus infi:ction resulted in a cessation of virus growth accompanied by a lack of development and possible regression in skein-like intranuclear inclusions together with a depletion in cytoplasmic dense bodies. Such changes were accompanied by the appearance of nuclear dense bodies. These were also present when virus growth was reduced (5 gM-DHPG) rather than completely inhibited (50 gM-DHPG) by treatment initiated from the time of infection. The nuclear bodies were predominantly of a reticular type structure after the early treatment but mainly of a homogeneous form when virus growth was interrupted at late times. Their presence appeared to be connected with the ability of infected cells to initiate the synthesis of late proteins and their morphology may relate to the extent of such protein synthesis. Unlike cytoplasmic dense bodies, provisional findings on the characterization of the nuclear bodies suggested that the 691( matrix protein was not present in abundance.

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Section: Methodsmentioning

confidence: 99%

Fine Structure of Cells Infected with Human Cytomegalovirus After Treatment with 9-(1,3-Dihydroxy-2-propoxymethyl)guanine

Mobberley¹,

Ryder²,

Hart³

et al. 1987

Journal of General Virology

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“…Infected and uninfected cells were exposed to [35S]methionine (10 IxCi/ml) for 2 h at selected times, harvested and stored frozen before analysis by discontinuous SDS-PAGE (Tyms et al, 1984b). Polypeptides were separated in l0 % acrylamide gels and tranferred electrophoretically to nitrocellulose (Hybond, Amersham), stained with 0.1 ~ amido black and subjected to autoradiography (Fuji RX film).…”

Section: Cellsmentioning

confidence: 99%

Synthesis of Cytomegalovirus DNA Is an Antiviral Target Late in Virus Growth

Tyms

Davis

Clarke

et al. 1987

Journal of General Virology

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SUMMARYThe mechanism of action of %(1,3-dihydroxypropoxymethyl)guanine (DHPG) and phosphonoformic acid (PFA) but not 5-fluorouridinedeoxyribose (FUdR), provides selective action against cytomegalovirus (CMV)-coded events and this was used to demonstrate that the synthesis of viral DNA was continuous during the extended phase of virus growth. The synthesis de novo of viral DNA was measured by restriction enzyme analysis after exposure to [32p]orthophosphate and its interruption by DHPG or PFA resulted in a cessation in the extrusion of infective virus from treated cells. The rate of decline in infectivity appeared to correspond to the failure of cells to maintain the synthesis of late proteins once DNA synthesis was blocked. Thus, regulation of late protein synthesis appeared to be linked to synthesis de novo of viral DNA even at late stages in CMV growth. The synthesis of the polyamines spermidine and spermine, considered obligatory for CMV growth, was unaffected by early or late inhibition of viral DNA and this showed that some virus-induced events were unaffected by the restriction on virus growth by DHPG. This provided evidence that polyamine biosynthesis was a target independent of viral DNA synthesis per se, which may be important in future considerations of combined drug therapies.

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“…DNA analysis was also used to confirm the identity of prototype CMV strains Towne and Kerr and HSV type 2 strain WF. Details of the method used for restriction enzyme analysis are published elsewhere [11]. Briefly, virus-infected cells were exposed to [32p]orthophosphate, DNA was extracted and restriction enzyme fragments separated by agarose gel electrophoresis.…”

Section: Restriction Enzyme Analysismentioning

confidence: 99%

The susceptibility of adenovirus infection to the anti-cytomegalovirus drug, ganciclovir (DHPG)

Taylor

Jeffries

Taylor-Robinson³

et al. 1988

FEMS Microbiology Letters

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The nucleoside analogue ganciclovir has clinical efficacy in the treatment of serious infections with human cytomegalovirus (CMV) in AIDS patients. The mechanism of action of the drug against CMV is different from that described for herpes simplex viruses (HSVs) as the crucial formation of the monophosphate derivative appears to be carried out by cellular rather than virus‐coded enzymes. Adenovirus infections also induce the expression of cellular genes including kinase activity and a novel DNA polymerase and the results reported here show that these viruses are sensitive to ganciclovir. The 50% effective dose (ED50) range for known serotypes and one clinical isolate was 4.5−33 μM. By comparison with the sensitivity of CMV in vitro and the known clinical response of infections with this virus to ganciclovir, our results suggest that this drug or its analogous may form the basis of chemotherapy for adenovirus infections.

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Human Cytomegalovirus Infections in vitro after Treatment with Arildone

Cited by 10 publications

References 37 publications

Fine Structure of Cells Infected with Human Cytomegalovirus After Treatment with 9-(1,3-Dihydroxy-2-propoxymethyl)guanine

Fine Structure of Cells Infected with Human Cytomegalovirus After Treatment with 9-(1,3-Dihydroxy-2-propoxymethyl)guanine

Synthesis of Cytomegalovirus DNA Is an Antiviral Target Late in Virus Growth

The susceptibility of adenovirus infection to the anti-cytomegalovirus drug, ganciclovir (DHPG)

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