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Heterocellular induction of interferon by negative-sense RNA viruses.

Chen S, Short JA, Young DF, Killip MJ, Schneider M, Goodbourn S, Randall RE - Virology (2010)

Bottom Line: The infection of cells by RNA viruses is associated with the recognition of virus PAMPs (pathogen-associated molecular patterns) and the production of type I interferon (IFN).Here we present data on the dynamics of IFN production and response during developing infections by paramyxoviruses, influenza A virus and bunyamwera virus.We show that only a limited number of infected cells are responsible for the production of IFN, and that this heterocellular production is a feature of the infecting virus as opposed to an intrinsic property of the cells.

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Centre for Biomolecular Sciences, BMS Building, North Haugh, University of St. Andrews, St. Andrews, Fife, KY16 9ST, UK.

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Detection of GFP-positive cells following infection of A549/pr(IFN-β).GFP cells with a panel of negative strand RNA viruses. A549/pr(IFN-β).GFP cells were infected with MuV (cl3), PIV2, PIV3, PIV5, FLUAV or BUNV at 2–5 pfu/cell. At 16 h p.i. the cells were fixed and immunostained with an anti-NP mAb. GFP-positive and virus-infected cells were visualised by fluorescence microscopy. The presence of the nuclei in the merge images was visualised by DAPI staining. Note that MuV (cl3/30) was originally plaque-purified from MuV (ori).
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f0015: Detection of GFP-positive cells following infection of A549/pr(IFN-β).GFP cells with a panel of negative strand RNA viruses. A549/pr(IFN-β).GFP cells were infected with MuV (cl3), PIV2, PIV3, PIV5, FLUAV or BUNV at 2–5 pfu/cell. At 16 h p.i. the cells were fixed and immunostained with an anti-NP mAb. GFP-positive and virus-infected cells were visualised by fluorescence microscopy. The presence of the nuclei in the merge images was visualised by DAPI staining. Note that MuV (cl3/30) was originally plaque-purified from MuV (ori).

Mentions: The data above show that efficient inducers of IFN-β (i.e. a MuV preparation rich in DI particles, or synthetic PAMPs; data not shown) cause a robust induction of GFP expression in the majority of A549/pr(IFN-β).GFP cells, with responsive cells producing similar levels of fluorescence. We next used the A549/pr(IFN-β).GFP cells to monitor the activation of the IFN-β promoter in individual cells during infection by viruses generated by passage at low m.o.i. Such preparations would be typical working stocks, lacking or low in DI particles, and are generally poor inducers of IFN-β (see for example Poole et al., 2002). Strikingly, in contrast to the extensive induction seen by the DI-rich preparation, MuV(ori) — see Fig. 2, a plaque-purified preparation of MuV, termed MuV cl3/30 (Young et al., 2009), showed high infectivity but only generated a few GFP-positive cells (Fig. 3). Each of the GFP-positive cells showed a similar degree of fluorescence, as described above for the MuV(ori) infection.


Heterocellular induction of interferon by negative-sense RNA viruses.

Chen S, Short JA, Young DF, Killip MJ, Schneider M, Goodbourn S, Randall RE - Virology (2010)

Detection of GFP-positive cells following infection of A549/pr(IFN-β).GFP cells with a panel of negative strand RNA viruses. A549/pr(IFN-β).GFP cells were infected with MuV (cl3), PIV2, PIV3, PIV5, FLUAV or BUNV at 2–5 pfu/cell. At 16 h p.i. the cells were fixed and immunostained with an anti-NP mAb. GFP-positive and virus-infected cells were visualised by fluorescence microscopy. The presence of the nuclei in the merge images was visualised by DAPI staining. Note that MuV (cl3/30) was originally plaque-purified from MuV (ori).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2963793&req=5

f0015: Detection of GFP-positive cells following infection of A549/pr(IFN-β).GFP cells with a panel of negative strand RNA viruses. A549/pr(IFN-β).GFP cells were infected with MuV (cl3), PIV2, PIV3, PIV5, FLUAV or BUNV at 2–5 pfu/cell. At 16 h p.i. the cells were fixed and immunostained with an anti-NP mAb. GFP-positive and virus-infected cells were visualised by fluorescence microscopy. The presence of the nuclei in the merge images was visualised by DAPI staining. Note that MuV (cl3/30) was originally plaque-purified from MuV (ori).
Mentions: The data above show that efficient inducers of IFN-β (i.e. a MuV preparation rich in DI particles, or synthetic PAMPs; data not shown) cause a robust induction of GFP expression in the majority of A549/pr(IFN-β).GFP cells, with responsive cells producing similar levels of fluorescence. We next used the A549/pr(IFN-β).GFP cells to monitor the activation of the IFN-β promoter in individual cells during infection by viruses generated by passage at low m.o.i. Such preparations would be typical working stocks, lacking or low in DI particles, and are generally poor inducers of IFN-β (see for example Poole et al., 2002). Strikingly, in contrast to the extensive induction seen by the DI-rich preparation, MuV(ori) — see Fig. 2, a plaque-purified preparation of MuV, termed MuV cl3/30 (Young et al., 2009), showed high infectivity but only generated a few GFP-positive cells (Fig. 3). Each of the GFP-positive cells showed a similar degree of fluorescence, as described above for the MuV(ori) infection.

Bottom Line: The infection of cells by RNA viruses is associated with the recognition of virus PAMPs (pathogen-associated molecular patterns) and the production of type I interferon (IFN).Here we present data on the dynamics of IFN production and response during developing infections by paramyxoviruses, influenza A virus and bunyamwera virus.We show that only a limited number of infected cells are responsible for the production of IFN, and that this heterocellular production is a feature of the infecting virus as opposed to an intrinsic property of the cells.

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Centre for Biomolecular Sciences, BMS Building, North Haugh, University of St. Andrews, St. Andrews, Fife, KY16 9ST, UK.

Show MeSH
Related in: MedlinePlus