PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins
Identifieur interne : 000C45 ( Main/Curation ); précédent : 000C44; suivant : 000C46PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins
Auteurs : Meshal Beidas [Koweït] ; Wassim Chehadeh [Koweït]Source :
- Open Forum Infectious Diseases [ 2328-8957 ] ; 2017.
Abstract
Human coronavirus OC43 (HCoV-OC43) causes common cold, and is associated with severe respiratory symptoms in infants, elderly and immunocompromised patients. HCoV-OC43 is a member of
HCoV-OC43 membrane (M), nucleocapsid (N), ns5a and ns2a mRNA were amplified and cloned into the pAcGFP1-N expression vector (Clontech), followed by transfection in HEK-293 cells. Expression of M, N, ns5a and ns2a proteins were confirmed by indirect immunofluorescence test. Three days post-transfection, the cells were challenged by Sendai virus. The Human Antiviral Response PCR array system (Qiagen) was used to profile the antiviral gene expression in HEK-293 cells, using the fold regulation comparison and the manual normalisation methods.
Around 50–60 genes were downregulated by HCoV-OC43 proteins, the most prominent genes being those critical for the activation of transcription factors involved in the antiviral response like interferon regulatory factors (IRFs) and activator protein 1 (AP-1). Among the most important downregulated genes were those coding for Interferons (IFNs) mitogen-activated protein kinases (MAPKs), pro-apoptotic and pyroptotic proteins (Caspases, cathepsins, tumour necrosis factor), pro-inflammatory cytokines (Interleukins), pattern recognition receptors (PRRs; toll-like receptors and NOD-like receptors) and their signaling transduction proteins (TICAM1, MAVS).
This study shows for the first time that similarly to SARS-CoV and MERS-CoV, HCoV-OC43 has the ability to downregulate the transcription of genes critical for the activation of different antiviral signaling pathways.
Url:
DOI: 10.1093/ofid/ofx163.728
PubMed: NONE
PubMed Central: 7107054
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PMC:7107054Le document en format XML
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<author><name sortKey="Beidas, Meshal" sort="Beidas, Meshal" uniqKey="Beidas M" first="Meshal" last="Beidas">Meshal Beidas</name>
<affiliation wicri:level="1"><nlm:aff id="AF0001"><institution>Microbiology, Kuwait University</institution>
,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
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<country xml:lang="fr">Koweït</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<author><name sortKey="Chehadeh, Wassim" sort="Chehadeh, Wassim" uniqKey="Chehadeh W" first="Wassim" last="Chehadeh">Wassim Chehadeh</name>
<affiliation wicri:level="1"><nlm:aff id="AF0001"><institution>Microbiology, Kuwait University</institution>
,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
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<country xml:lang="fr">Koweït</country>
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<author><name sortKey="Beidas, Meshal" sort="Beidas, Meshal" uniqKey="Beidas M" first="Meshal" last="Beidas">Meshal Beidas</name>
<affiliation wicri:level="1"><nlm:aff id="AF0001"><institution>Microbiology, Kuwait University</institution>
,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
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<author><name sortKey="Chehadeh, Wassim" sort="Chehadeh, Wassim" uniqKey="Chehadeh W" first="Wassim" last="Chehadeh">Wassim Chehadeh</name>
<affiliation wicri:level="1"><nlm:aff id="AF0001"><institution>Microbiology, Kuwait University</institution>
,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
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<series><title level="j">Open Forum Infectious Diseases</title>
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<front><div type="abstract" xml:lang="en"><title>Abstract</title>
<sec id="s1"><title>Background</title>
<p>Human coronavirus OC43 (HCoV-OC43) causes common cold, and is associated with severe respiratory symptoms in infants, elderly and immunocompromised patients. HCoV-OC43 is a member of <italic>Betacoronavirus</italic>
genus that includes also the Severe Acute Respiratory Syndrome (SARS) and the Middle East Respiratory Syndrome (MERS) coronaviruses. Both SARS-CoV and MERS-CoV were shown to express proteins with the potential to evade early innate immune responses. However, the ability of HCoV-OC43 to antagonise the intracellular antiviral defences has not yet been investigated. The objective of this study was to investigate the role of HCoV-OC43 structural (membrane and nucleocapsid) and accessory (ns5a and ns2a) proteins in the modulation of antiviral gene expression profile in human embryonic kidney 293 (HEK-293) cells using PCR array analysis.</p>
</sec>
<sec id="s2"><title>Methods</title>
<p>HCoV-OC43 membrane (M), nucleocapsid (N), ns5a and ns2a mRNA were amplified and cloned into the pAcGFP1-N expression vector (Clontech), followed by transfection in HEK-293 cells. Expression of M, N, ns5a and ns2a proteins were confirmed by indirect immunofluorescence test. Three days post-transfection, the cells were challenged by Sendai virus. The Human Antiviral Response PCR array system (Qiagen) was used to profile the antiviral gene expression in HEK-293 cells, using the fold regulation comparison and the manual normalisation methods.</p>
</sec>
<sec id="s3"><title>Results</title>
<p>Around 50–60 genes were downregulated by HCoV-OC43 proteins, the most prominent genes being those critical for the activation of transcription factors involved in the antiviral response like interferon regulatory factors (IRFs) and activator protein 1 (AP-1). Among the most important downregulated genes were those coding for Interferons (IFNs) mitogen-activated protein kinases (MAPKs), pro-apoptotic and pyroptotic proteins (Caspases, cathepsins, tumour necrosis factor), pro-inflammatory cytokines (Interleukins), pattern recognition receptors (PRRs; toll-like receptors and NOD-like receptors) and their signaling transduction proteins (TICAM1, MAVS).</p>
</sec>
<sec id="s4"><title>Conclusion</title>
<p>This study shows for the first time that similarly to SARS-CoV and MERS-CoV, HCoV-OC43 has the ability to downregulate the transcription of genes critical for the activation of different antiviral signaling pathways.</p>
</sec>
<sec id="s5"><title>Disclosures</title>
<p><bold>All authors:</bold>
No reported disclosures.</p>
</sec>
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