A simple and rapid microassay for the titration of human respiratory syncytial virus

Trépanier, Pierre; Payment, Pierre; Trudel, Michel

doi:10.1016/0166-0934(80)90051-8

Cited by 11 publications

(10 citation statements)

References 11 publications

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“…We also modified an RSV syncytia-formation assay (Lambert et al, 1980;Trepanier et al, 1980) in order to measure the amounts of infectious RSV that either remained associated with the infected cells or had been secreted into the medium and recorded as 'cell-free'. Ceil-associated virus was released by a freeze-thaw, sonication or osmotic lysis procedure.…”

Section: Inhibition Of Infectious Virusmentioning

confidence: 99%

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Antiviral Activities of Peptides with Sequences Related to the Cytoplasmic Domain of the Respiratory Syncytial Virus Glycoproteins

Ryan

Schlesinger

1995

Antivir Chem Chemother

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SummaryA set of peptides from 6-10 amino acids in length with sequences corresponding to the respiratory syncytial virus (RSV) attachment glycoprotein, G, has been found to inhibit secretion of virus particles from two human cell lines infected with RSV. Peptides of similar lengths with sequences related to the fusion (F) protein were not effective. A modified assay for syncytia formation was used to measure infectious particles and we found that cell-free infectious RSV was inhibited by those same peptides that blocked particle release. However, only about 5-20% of the total infectious virus formed in the cultured cells was released from the cells -the remaining was assayable only after lysis of the infected cells. Formation of this major fraction of infectious, ceIl-associated RSV was not affected by the peptides. The inhibitory peptides were ineffective in blocking release of infectious virus from cells infected with influenza, vesicular stomatitis and Semliki Forest viruses. Only a set of specific amino acids in these peptides were inhibitory and they consisted of a very hydrophobic sequence that also required a cysteine residue.

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Section: Inhibition Of Infectious Virusmentioning

confidence: 99%

“…To measure infectious RSV, the assay of syncytia formation as described by Lambert et al, (1980) and Trepanier et al, (1980) was modified. Virus stocks were titered using 1:10 serial dilutions in PBS plus 1% HI-FBS.…”

Section: Assays For Virus Formationmentioning

confidence: 99%

Antiviral Activities of Peptides with Sequences Related to the Cytoplasmic Domain of the Respiratory Syncytial Virus Glycoproteins

Ryan

Schlesinger

1995

Antivir Chem Chemother

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“…More rapid systems have also been described, in which syncytia or plaques can be microscopically enumerated in tissue culture or under a semi-fluid overlay of methylcellulose (4) in as few as 3 days. A microassay using a 3-7-day incubation period has also been described (10). We report the development of a simple and rapid titration assay for RSV using a quantitative shell vial approach.…”

mentioning

confidence: 99%

“…Since the isolation of human RSV by Chanock et al in 1957 (3), investigators have relied on endpoint dilution or conventional plaque assay methods for its titration. The latter is considered to be more accurate (10). In RSV plaque assays, plaques that can be macroscopically counted in 9 days under agar overlays (7) or in 13 days under a liquid overlay (8) give reliable and reproducible titrations of a stock aliquot of virus.…”

mentioning

confidence: 99%

Overnight Titration of Human Respiratory Syncytial Virus Using Quantitative Shell Vial Amplification

Domachowske

Bonville

1998

BioTechniques

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“…The plate (without cover) was placed on ice and illuminated with UV-C at a height of 10 cm for 30 minutes. Loss of virus titer was verified by syncytia titration on vero cells (Trepanier et al, 1980).…”

Section: Respiratory Syncytial Virusmentioning

confidence: 99%

Augmenting antiviral host defense in the respiratory epithelium

Fischer¹

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The airway epithelium has many roles in innate immunity including detection of pathogens and transmitting danger signals to other cell types. However, its role as a primary defender against infection is not well recognized. We have investigated methods of augmenting antiviral immunity by application of agents that stimulate viral killing, either in the extracellular space or within the cytoplasm. A recently described property of airway epithelial cells is direct oxidative killing of bacteria through the coordination of Duox and lactoperoxidase enzymes. We have exploited this property by supplementing airway cells with the lactoperoxidase substrate iodide to prevent viral infection. A second method for enhancing antiviral defenses is to supply small interfering RNAs (siRNAs) targeting essential viral genes. We have optimized antiviral siRNAs targeting respiratory syncytial virus by designing them to specifically target positive sense viral RNAs. Finally, we have initiated a project to discover host defense genes that are expressed in either the submucosal glands surface epithelium of human airway. This information will enable a better characterization of the roles for these structures in host defense pathways, and may identify other targets for augmentation of antiviral immunity.Abstract Approved: __________________________________ Thesis Supervisor __________________________________

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A simple and rapid microassay for the titration of human respiratory syncytial virus

Cited by 11 publications

References 11 publications

Antiviral Activities of Peptides with Sequences Related to the Cytoplasmic Domain of the Respiratory Syncytial Virus Glycoproteins

Antiviral Activities of Peptides with Sequences Related to the Cytoplasmic Domain of the Respiratory Syncytial Virus Glycoproteins

Overnight Titration of Human Respiratory Syncytial Virus Using Quantitative Shell Vial Amplification

Augmenting antiviral host defense in the respiratory epithelium

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