Suitability of infection of cells in suspension for detection of herpes simplex virus

Luker, Gerri; Chow, Connie; Richards, D F; Johnson, F. Brent

doi:10.1128/jcm.29.7.1554-1557.1991

Cited by 11 publications

(3 citation statements)

References 24 publications

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“…Infectivity assays on the supernatants were performed in flat-sided tissue culture tubes (Nunc), as described previously (Johnson et al, 2004), to detect infectious virus particles released into the medium of the cultures. Following incubation, cells were fixed with formaldehyde/ethanol/acetic acid and stained with an immunoperoxidase (IP) stain to identify cells positive for viral antigen (Luker et al, 1991). The stained cells were counted and infectious virus titres determined and notated as focus-forming units (f.f.u.)…”

Section: Methodsmentioning

confidence: 99%

Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis

Abdel-Latif

Murray

Renberg

et al. 2006

Journal of General Virology

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Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis The helper-independent bovine parvovirus (BPV) was studied to determine its effect on host embryonic bovine tracheal (EBTr) cells: whether the ultimate outcome of infection results in apoptotic cell death or cell death by necrosis. Infected cells were observed for changes marking apoptosis. Observations of alterations in nuclear morphology, membrane changes, apoptotic body formation, membrane phosphatidylserine inversions, caspase activation and cell DNA laddering in infected cells were not indicative of apoptosis. On the other hand, at the end of the virus replication cycle, infected cells released viral haemagglutinin and infectious virus particles, as would be expected from cell membrane failure. Moreover, the infected cells released lactate dehydrogenase (LDH), release of which is a marker of necrosis. LDH release into the cell medium correlated directly with viral m.o.i. and time post-infection. Furthermore, assessment of mitochondrial dehydrogenase activity was consistent with cell death by necrosis. Taken together, these findings indicate that cell death in BPV-infected EBTr cells is due to necrosis, as defined by infected-cell membrane failure and release of the cell contents into the extracellular environment.

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Section: Methodsmentioning

confidence: 99%

Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis

Abdel-Latif

Murray

Renberg

et al. 2006

Journal of General Virology

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“…Cell monolayers of C1008 cells in 24-well culture plates (Corning) were exposed to drug at varying concentrations, infected with HHV-2, strain G (ATCC VR-734), at a multiplicity of infection (MOI) 0.001, incubated at 36 °C, then processed by fixation and immunoperoxidase staining (Luker et al. 1991 ). The plaques were observed microscopically and counted.…”

Section: Methodsmentioning

confidence: 99%

Isolation and identification of compounds from Kalanchoe pinnata having human alphaherpesvirus and vaccinia virus antiviral activity

Cryer

Lane

Greer

et al. 2017

Pharmaceutical Biology

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Context:Kalanchoe pinnata (Lam.) Pers. (Crassulaceae) is a succulent plant that is known for its traditional antivirus and antibacterial usage.Objective: This work examines two compounds identified from the K. pinnata plant for their antivirus activity against human alphaherpesvirus (HHV) 1 and 2 and vaccinia virus (VACV).Materials and methods: Compounds KPB-100 and KPB-200 were isolated using HPLC and were identified using NMR and MS. Both compounds were tested in plaque reduction assay of HHV-2 wild type (WT) and VACV. Both compounds were then tested in virus spread inhibition and virus yield reduction (VYR) assays of VACV. KPB-100 was further tested in viral cytopathic effect (CPE) inhibition assay of HHV-2 TK-mutant and VYR assay of HHV-1 WT.Results: KPB-100 and KPB-200 inhibited HHV-2 at IC50 values of 2.5 and 2.9 μg/mL, respectively, and VACV at IC50 values of 3.1 and 7.4 μg/mL, respectively, in plaque reduction assays. In virus spread inhibition assay of VACV KPB-100 and KPB-200 yielded IC50 values of 1.63 and 13.2 μg/mL, respectively, and KPB-100 showed a nearly 2-log reduction in virus in VYR assay of VACV at 20 μg/mL. Finally, KPB-100 inhibited HHV-2 TK- at an IC50 value of 4.5 μg/mL in CPE inhibition assay and HHV-1 at an IC90 of 3.0 μg/mL in VYR assay.Discussion and conclusion: Both compounds are promising targets for synthetic optimization and in vivo study. KPB-100 in particular showed strong inhibition of all viruses tested.

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“…DMEM containing 2 % FBS was added and the cultures were incubated at 36 uC for 2 days. The monolayers were fixed in formaldehyde/ethanol/acetic acid (FAA) fixative (Luker et al, 1991) and stained with guinea pig anti-BPV antibody using an immunoperoxidase method described previously (Luker et al, 1991), with the following modification: the chromogen consisted of 20 mg 4-chloro-1-naphthol in 100 ml of a solution composed of 2 ml DMSO, 10 ml 95 % ethanol, 0?1 ml 30 % hydrogen peroxide and 87?9 ml distilled water.…”

Section: Some Inhibition Tests Were Carried Out In Madin-darby Bovinementioning

confidence: 99%

Attachment of bovine parvovirus to sialic acids on bovine cell membranes

Johnson

Fenn

Owens

et al. 2004

Journal of General Virology

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Although it has previously been shown that bovine parvovirus (BPV) attaches to the sialated glycoprotein glycophorin A on erythrocytes, the nature of virus-binding moieties on mammalian nucleated cells is less clear. Buffalo lung fibroblasts (Bu), primary bovine embryonic kidney cells, Madin-Darby bovine kidney cells and bovine embryonic trachea (EBTr) cells were assessed for molecules capable of binding BPV. Competition studies were carried out on both erythrocyte and nucleated cell targets using a variety of sialated compounds and sialic acid-negative compounds. Glycophorin A was found to inhibit BPV binding, while mucin exhibited low-level inhibition. These two sialated compounds also blocked attachment of BPV-modified microsphere carriers to the Bu cell membrane. Influenza A virus was used as a sialic acid competitor and interfered with BPV attachment to erythrocytes and replication in Bu cells. Significantly, the enzyme sialidase removed BPV-binding sites from Bu and EBTr cells. The binding sites could be reconstituted on sialidase-treated cells by the enzymes a-2,3-O-sialyltransferase and a-2,3-N-sialyltransferase. These results indicated that BPV can attach to sialic acid on cell membranes and that the sialylglycoproteins available for virus attachment appear to contain both N-and O-linked carbohydrate moieties, but that not all members of the sialic acid family can bind BPV. Moreover, there may be other moieties that can bind BPV, which may act as either primary or secondary receptors.

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Suitability of infection of cells in suspension for detection of herpes simplex virus

Cited by 11 publications

References 24 publications

Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis

Cell death in bovine parvovirus-infected embryonic bovine tracheal cells is mediated by necrosis rather than apoptosis

Isolation and identification of compounds from Kalanchoe pinnata having human alphaherpesvirus and vaccinia virus antiviral activity

Attachment of bovine parvovirus to sialic acids on bovine cell membranes

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