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Influenza virus assays based on virus-inducible reporter cell lines.

Li Y, Larrimer A, Curtiss T, Kim J, Jones A, Baird-Tomlinson H, Pekosz A, Olivo PD - Influenza Other Respir Viruses (2009)

Bottom Line: A strategy for influenza A virus-induction of a reporter gene was recently described.Upon infection of the cells with influenza virus the virus-inducible reporter gene segment (VIRGS) is replicated and transcribed by the viral polymerase complex resulting in reporter gene expression.Reporter gene induction occurs after infection with a number of laboratory strains and clinical isolates of influenza virus including several H5N1 strains.

View Article: PubMed Central - PubMed

Affiliation: Diagnostic Hybrids Inc., Athens, OH, USA.

ABSTRACT

Background: Virus-inducible reporter genes have been used as the basis of virus detection and quantitation assays for a number of viruses. A strategy for influenza A virus-induction of a reporter gene was recently described. In this report, we describe the extension of this strategy to influenza B virus, the generation of stable cell lines with influenza A and B virus-inducible reporter genes, and the use of these cells in various clinically relevant viral assays. Each of the cell lines described herein constitutively express an RNA transcript that contains a reporter gene coding region flanked by viral 5¢- and 3¢-untranslated regions (UTR) and therefore mimics an influenza virus genomic segment. Upon infection of the cells with influenza virus the virus-inducible reporter gene segment (VIRGS) is replicated and transcribed by the viral polymerase complex resulting in reporter gene expression.

Findings: Reporter gene induction occurs after infection with a number of laboratory strains and clinical isolates of influenza virus including several H5N1 strains. The induction is dose-dependent and highly specific for influenza A or influenza B viruses.

Conclusions: These cell lines provide the basis of simple, rapid, and objective assays that involve virus quantitation such as determination of viral titer, assessment of antiviral susceptibility, and determination of antibody neutralization titer. These cell lines could be very useful for influenza virus researchers and vaccine manufacturers.

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Related in: MedlinePlus

 Time course of luciferase expression of ELVIRA® Flu A‐luc or Flu B‐Rluc cells following infection with influenza viruses. ELVIRA® Flu A‐luc or Flu B‐Rluc cells were infected with influenza A/Wisconsin67/05 or influenza B/Malaysia2506/04 at the indicated MOI. Luciferase activity was measured at various time points after virus infection and expressed as RLU/well. Results shown are mean of 3 replicates. (A) Luciferase activity of ELVIRA® Flu A‐luc cells infected with influenza A/Wisconsin67/05. The insert shows data from the early time points. (B) Luciferase activity of ELVIRA® Flu B‐Rluc cells infected with influenza B Malaysia2506/04.
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f1:  Time course of luciferase expression of ELVIRA® Flu A‐luc or Flu B‐Rluc cells following infection with influenza viruses. ELVIRA® Flu A‐luc or Flu B‐Rluc cells were infected with influenza A/Wisconsin67/05 or influenza B/Malaysia2506/04 at the indicated MOI. Luciferase activity was measured at various time points after virus infection and expressed as RLU/well. Results shown are mean of 3 replicates. (A) Luciferase activity of ELVIRA® Flu A‐luc cells infected with influenza A/Wisconsin67/05. The insert shows data from the early time points. (B) Luciferase activity of ELVIRA® Flu B‐Rluc cells infected with influenza B Malaysia2506/04.

Mentions: Virus‐inducible reporter gene expression from the ELVIRA® Flu A‐luc and ELVIRA® Flu B‐Rluc cell lines was assessed after infection with influenza A and B viruses respectively at high and low MOI. At various times after infection cells were lysed and luciferase activity was measured (Figure 1). The graphic display of the data reveals classic single and multistep viral growth kinetics and a time course very similar to infectious virus production on these cells. These results reveal that measurement of luciferase activity provides a valid surrogate marker of virus replication.


Influenza virus assays based on virus-inducible reporter cell lines.

Li Y, Larrimer A, Curtiss T, Kim J, Jones A, Baird-Tomlinson H, Pekosz A, Olivo PD - Influenza Other Respir Viruses (2009)

 Time course of luciferase expression of ELVIRA® Flu A‐luc or Flu B‐Rluc cells following infection with influenza viruses. ELVIRA® Flu A‐luc or Flu B‐Rluc cells were infected with influenza A/Wisconsin67/05 or influenza B/Malaysia2506/04 at the indicated MOI. Luciferase activity was measured at various time points after virus infection and expressed as RLU/well. Results shown are mean of 3 replicates. (A) Luciferase activity of ELVIRA® Flu A‐luc cells infected with influenza A/Wisconsin67/05. The insert shows data from the early time points. (B) Luciferase activity of ELVIRA® Flu B‐Rluc cells infected with influenza B Malaysia2506/04.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4940803&req=5

f1:  Time course of luciferase expression of ELVIRA® Flu A‐luc or Flu B‐Rluc cells following infection with influenza viruses. ELVIRA® Flu A‐luc or Flu B‐Rluc cells were infected with influenza A/Wisconsin67/05 or influenza B/Malaysia2506/04 at the indicated MOI. Luciferase activity was measured at various time points after virus infection and expressed as RLU/well. Results shown are mean of 3 replicates. (A) Luciferase activity of ELVIRA® Flu A‐luc cells infected with influenza A/Wisconsin67/05. The insert shows data from the early time points. (B) Luciferase activity of ELVIRA® Flu B‐Rluc cells infected with influenza B Malaysia2506/04.
Mentions: Virus‐inducible reporter gene expression from the ELVIRA® Flu A‐luc and ELVIRA® Flu B‐Rluc cell lines was assessed after infection with influenza A and B viruses respectively at high and low MOI. At various times after infection cells were lysed and luciferase activity was measured (Figure 1). The graphic display of the data reveals classic single and multistep viral growth kinetics and a time course very similar to infectious virus production on these cells. These results reveal that measurement of luciferase activity provides a valid surrogate marker of virus replication.

Bottom Line: A strategy for influenza A virus-induction of a reporter gene was recently described.Upon infection of the cells with influenza virus the virus-inducible reporter gene segment (VIRGS) is replicated and transcribed by the viral polymerase complex resulting in reporter gene expression.Reporter gene induction occurs after infection with a number of laboratory strains and clinical isolates of influenza virus including several H5N1 strains.

View Article: PubMed Central - PubMed

Affiliation: Diagnostic Hybrids Inc., Athens, OH, USA.

ABSTRACT

Background: Virus-inducible reporter genes have been used as the basis of virus detection and quantitation assays for a number of viruses. A strategy for influenza A virus-induction of a reporter gene was recently described. In this report, we describe the extension of this strategy to influenza B virus, the generation of stable cell lines with influenza A and B virus-inducible reporter genes, and the use of these cells in various clinically relevant viral assays. Each of the cell lines described herein constitutively express an RNA transcript that contains a reporter gene coding region flanked by viral 5¢- and 3¢-untranslated regions (UTR) and therefore mimics an influenza virus genomic segment. Upon infection of the cells with influenza virus the virus-inducible reporter gene segment (VIRGS) is replicated and transcribed by the viral polymerase complex resulting in reporter gene expression.

Findings: Reporter gene induction occurs after infection with a number of laboratory strains and clinical isolates of influenza virus including several H5N1 strains. The induction is dose-dependent and highly specific for influenza A or influenza B viruses.

Conclusions: These cell lines provide the basis of simple, rapid, and objective assays that involve virus quantitation such as determination of viral titer, assessment of antiviral susceptibility, and determination of antibody neutralization titer. These cell lines could be very useful for influenza virus researchers and vaccine manufacturers.

Show MeSH
Related in: MedlinePlus