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An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection
Published on: December 10, 2013
Effect of interferon-alpha on measles virus replication in human peripheral blood mononuclear cells
R Leopardi1, T Hyypiä, R Vainionpää
1Department of Virology, University of Turku, Finland.
Abstract:
We analyzed the effect of exogenous human leukocyte interferon (IFN)-alpha on measles virus (MV) replication in human peripheral blood mononuclear cells (PBMC). The release of infectious virus was progressively reduced by increasing concentrations of IFN-alpha, and blocked with an IFN-alpha concentration of 1000 U/ml. In order to detect a possible target of this inhibitory effect, viral transcription and translation events were analyzed. The synthesis of MV mRNAs was reduced, but not blocked, in the presence of IFN-alpha. However, this effect was not specific on the viral RNAs, but due to a general inhibition of RNA synthesis in IFN-treated PBMC. The expression of viral polypeptides was also inhibited in a dose-dependent manner by exogenous IFN-alpha, but a low level of protein synthesis was detected by both Western blotting and immunofluorescence techniques, even with the maximum amount of IFN-alpha used (1000 U/ml). These findings account for a partial maintenance of the viral replicative cycle, even when the production of infectious virus is blocked. Moreover, the effect of IFN-alpha is not specifically targeted on the virus macromolecular synthesis.
Insights
Exogenous human leukocyte interferon-alpha (IFN-alpha) inhibits measles virus (MV) replication in human cells. While IFN-alpha blocks infectious virus release, it broadly affects cellular RNA and protein synthesis, not specifically targeting viral macromolecules.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Measles virus (MV) is a significant human pathogen.
- Interferon-alpha (IFN-alpha) is a key antiviral cytokine.
- Understanding host-pathogen interactions is crucial for antiviral strategies.
Purpose of the Study:
- To investigate the effect of exogenous human leukocyte interferon-alpha (IFN-alpha) on measles virus (MV) replication.
- To determine the specific molecular targets of IFN-alpha's antiviral activity against MV in human peripheral blood mononuclear cells (PBMC).
Main Methods:
- Treatment of human PBMC with varying concentrations of IFN-alpha.
- Analysis of infectious MV release.
- Assessment of viral mRNA synthesis and overall RNA synthesis.
- Evaluation of viral polypeptide expression using Western blotting and immunofluorescence.
Main Results:
- IFN-alpha progressively reduced infectious MV release, completely blocking it at 1000 U/ml.
- Viral mRNA synthesis was reduced but not blocked by IFN-alpha.
- IFN-alpha caused a general inhibition of RNA synthesis in PBMC, not specific to viral RNA.
- Viral protein expression was dose-dependently inhibited, with low-level synthesis detected even at maximum IFN-alpha concentrations.
Conclusions:
- Exogenous IFN-alpha effectively inhibits infectious measles virus production in human cells.
- The antiviral effect of IFN-alpha is not specifically targeted at viral macromolecular synthesis but involves broad inhibition of host cell RNA and protein synthesis.
- A partial viral replicative cycle may persist despite the blockage of infectious virus production.
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