A nonlytic transforming mutant of herpes simplex virus type 2

Howett, Mary K.; Rapp, Fred

doi:10.1002/jmv.1890020106

Cited by 2 publications

(1 citation statement)

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“…were combined with 14C-labeled TC-7 cell DNA (8.5 pLl, -5,000 cpm), 5.0x STEP buffer (50 mM Tris hydrochloride [pH 7.51-40 mM EDTA), sodium dodecyl sulfate (0.5%), and proteinase K (2 mg/ml) to a final volume of 1 ml. Samples then were digested at 50°C for 1 h. After digestion, the 1-ml labeled sample was added to 3 ml of cesium chloride solution in 0.1 x SSC (1 x SSC is 0.15 M NaCl plus 0.015 M sodium citrate) (final density of gradient, 1.71 g/ml) and centrifuged at 33,000 rpm for 60 h with a 40.30 fixed-angle rotor (19). After centrifugation, 8-drop fractions (-0.1 ml) were collected and 50-pul samples were assayed for trichloroacetic acid-precipitable counts on Whatman filter paper disks.…”

Section: Methodsmentioning

confidence: 99%

A human cytomegalovirus function inhibits replication of herpes simplex virus

Cockley

Shiraki

Rapp

1988

J Virol

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Human embryonic lung (HEL) cells infected with human cytomegalovirus (HCMV) restricted the replication of herpes simplex virus type 1 (HSV-1). A delay in HSV replication of 15 h as well as a consistent, almost 3 log inhibition of HSV replication in HCMV-infected cell cultures harvested 24 to 72 h after superinfection were observed compared with controls infected with HSV alone. Treatment of HCMV-infected HEL cells with cycloheximide (100 ,ug/mI) for 3 or 24 h, conditions known to result in accumulation of HCMV immediate-early and early mRNA, was demonstrated effective in blocking HCMV protein synthesis, as shown by immunoprecipitation with HCMV antibody-positive polyvalent serum. Cycloheximide treatment of HCMV-infected HEL cells and removal of the cycloheximide block before superinfection inhibited HSV-1 replication more efficiently than non-drug-treated superinfected controls. HCMV DNA-negative temperature-sensitive mutants restricted HSV as efficiently as wild-type HCMV suggesting that immediate-early and/or early events which occur before viral DNA synthesis are sufficient for inhibition of HSV. Inhibition of HSV-1 in HCMV-infected HEL cells was unaffected by elevated temperature (40.5°C). However, prior UV irradiation of HCMV removed the block to HSV replication, demonstrating the requirement for an active HCMV genome. HSV-2 replication was similarly inhibited in HCMV-infected HEL cells. However, replication of adenovirus, another DNA virus, was not restricted in these cells under the same conditions. Superinfection of HCMV-infected HEL cells with HSV-1 labeled with [3H]thymidine provided evidence that the labeled virus could penetrate to the nucleus of cells after superinfection. Evidence for penetration of superinfecting HSV into HCMV-infected cells was also provided by blot hybridization of HSV DNA synthesized in cells infected with HSV alone versus superinfected cell cultures at 0 and 48 h after superinfection. In addition, superinfection with vesicular stomatitis virus ruled out a role for interferon in restriction of HSV replication in this system.

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Section: Methodsmentioning

confidence: 99%