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Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
Published on: May 15, 2014
Comparative transcriptome analyses indicate enhanced cellular protection against FMDV in PK15 cells pretreated with
Yin Fu1, Zesen Zhu1, Huiyun Chang2
1State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410078, China.
Abstract:
Interferon gamma (IFN-γ) can induce a host antiviral response to foot and mouth disease virus (FMDV) in vivo and in vitro. To elucidate the mechanism of IFN-γ anti FMDV infection in host cells, high-throughput RNA sequencing was analyzed for systemic changes in gene expression profiles in PK15 cells infected by FMDV with or without IFN-γ pretreatment. More than 25 million reads, covering 1.2-1.5 Gb, were analyzed from each experiment panel. FMDV challenge altered the transcription of genes involved in positively and negatively regulating cell death or apoptosis; however, the expected immune suppression response was not obvious. IFN-γ pretreatment combined with FMDV infection normalized the increase in apoptosis. Furthermore, the transcription factors required for IFN-γ functioning, STAT1 and IRF1 were up-regulated by IFN-γ pretreatment and stimulated downstream IFN-stimulated genes (ISGs). These induced ISGs are mainly responsible for antigen processing, antigen presentation or antiviral defense. Interestingly, a synergistic effect on some ISGs, including OAS1, OAS2, MX1, MX2, RIG-I and IFIT1, was observed in the combined treatment compared to the IFN-γ treatment alone. The suggested effects identified by RNA sequencing were consistent with cellular morphology changes and confirmed by related protein markers. This is the first report exploring transcriptome alterations introduced by FMDV infection with or without IFN-γ pretreatment. The identified key host genes that control cell survival in vitro broaden our comprehensive understanding of how IFN-γ inhibits FMDV infection and may shed light on developing improved FMD control approaches.
Insights
Interferon gamma (IFN-γ) pretreatment helps control foot and mouth disease virus (FMDV) by normalizing apoptosis and upregulating antiviral genes. This study reveals key host genes involved in IFN-γ
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Foot and mouth disease virus (FMDV) poses a significant threat to livestock.
- Interferon gamma (IFN-γ) is known to induce antiviral responses against FMDV.
- The precise mechanisms of IFN-γ's anti-FMDV action in host cells require further elucidation.
Purpose of the Study:
- To investigate the molecular mechanisms by which IFN-γ inhibits FMDV infection in host cells.
- To analyze systemic gene expression changes in PK15 cells upon FMDV infection with or without IFN-γ pretreatment using high-throughput RNA sequencing.
Main Methods:
- High-throughput RNA sequencing of PK15 cells infected with FMDV, with and without prior IFN-γ treatment.
- Analysis of over 25 million reads (1.2-1.5 Gb) per sample to assess global gene expression profiles.
- Validation of RNA sequencing findings through cellular morphology assessments and protein marker analysis.
Main Results:
- FMDV infection altered genes related to apoptosis; IFN-γ pretreatment normalized FMDV-induced apoptosis.
- IFN-γ pretreatment upregulated STAT1 and IRF1, which in turn stimulated downstream IFN-stimulated genes (ISGs).
- A synergistic effect was observed on specific ISGs (OAS1, OAS2, MX1, MX2, RIG-I, IFIT1) with combined IFN-γ and FMDV treatment.
Conclusions:
- IFN-γ pretreatment modulates host gene expression to enhance antiviral defense against FMDV.
- The study identifies key host genes, including ISGs, that are crucial for controlling FMDV infection and cell survival.
- These findings provide a comprehensive understanding of IFN-γ's role in combating FMDV and may inform improved FMD control strategies.
