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Differential expression of interferon alpha and beta induced with Newcastle disease virus in mouse macrophage
A Hoss1, E C Zwarthoff, R Zawatzky
1Institute of Virus Research, German Cancer Research Center, Heidelberg.
Abstract:
Stimulation of mouse macrophages with Newcastle disease virus (NDV) leads to a rapid and high interferon (IFN) response. The magnitude of this response is influenced by the mouse genotype. We have analysed NDV-induced IFN production at the protein and mRNA levels in two different populations of macrophages derived from 'high producer' C57BL/6 and 'low producer' BALB/c mice in vitro. The data indicate that bone marrow and peritoneal macrophages from both strains grown in the presence of L cell conditioned medium (CM) as a source of macrophage colony-stimulating factor 1 (M-CSF) or purified murine M-CSF produce 10- to 50-fold more IFN on a per cell basis than cultures of resident peritoneal macrophages. These differences were also found when steady state levels of IFN mRNA were analysed. Differential analysis for the ratios of IFN-alpha and IFN-beta showed that CM- or M-CSF-cultured macrophages produced equal amounts of both IFN species as determined by specific monoclonal antibodies and hybridization experiments using IFN-alpha and IFN-beta DNA probes, whereas resident peritoneal macrophages induced under identical conditions produced almost exclusively IFN-beta. This suggests a stimulating effect of M-CSF on IFN synthesis in NDV-induced cultures of mouse macrophages, which is in part due to additional activation of IFN-alpha gene expression.
Insights
Newcastle disease virus (NDV) triggers a strong interferon (IFN) response in mouse macrophages. Macrophage colony-stimulating factor 1 (M-CSF) significantly boosts IFN production, particularly IFN-alpha, in cultured macrophages.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Newcastle disease virus (NDV) induces a robust interferon (IFN) response in macrophages.
- Mouse genotype influences the magnitude of this IFN response.
- Previous studies suggest macrophage activation status impacts IFN production.
Purpose of the Study:
- To investigate the effect of macrophage colony-stimulating factor 1 (M-CSF) on NDV-induced IFN production in vitro.
- To compare IFN production at protein and mRNA levels in different macrophage populations.
- To elucidate the role of M-CSF in regulating IFN-alpha and IFN-beta gene expression.
Main Methods:
- Primary macrophages were derived from C57BL/6 ('high producer') and BALB/c ('low producer') mice.
- Macrophages were cultured with or without L cell conditioned medium (CM) or purified murine M-CSF.
- NDV stimulation was used to induce IFN production.
- IFN protein levels were measured using specific monoclonal antibodies.
- IFN mRNA levels were analyzed using hybridization experiments with DNA probes.
Main Results:
- Macrophages cultured with CM or M-CSF produced 10- to 50-fold more IFN per cell than resident peritoneal macrophages.
- This enhancement was observed at both protein and mRNA levels.
- CM- or M-CSF-cultured macrophages produced both IFN-alpha and IFN-beta, while resident macrophages produced almost exclusively IFN-beta.
- M-CSF significantly increased IFN-alpha gene expression.
Conclusions:
- M-CSF significantly enhances NDV-induced IFN synthesis in mouse macrophages.
- This enhancement is partly due to M-CSF's ability to activate IFN-alpha gene expression.
- Macrophage activation status, influenced by M-CSF, plays a critical role in determining the type and quantity of IFN produced.