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
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins</title><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></nlm:aff><country xml:lang="fr">Koweït</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><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></nlm:aff><country xml:lang="fr">Koweït</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmc">7107054</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7107054</idno><idno type="RBID">PMC:7107054</idno><idno type="doi">10.1093/ofid/ofx163.728</idno><idno type="pmid">NONE</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">000636</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000636</idno><idno type="wicri:Area/Pmc/Curation">000636</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000636</idno><idno type="wicri:Area/Pmc/Checkpoint">000754</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000754</idno><idno type="wicri:Area/Ncbi/Merge">002C99</idno><idno type="wicri:Area/Ncbi/Curation">002C99</idno><idno type="wicri:Area/Ncbi/Checkpoint">002C99</idno><idno type="wicri:Area/Main/Merge">000C48</idno><idno type="wicri:Area/Main/Curation">000C45</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins</title><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></nlm:aff><country xml:lang="fr">Koweït</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><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></nlm:aff><country xml:lang="fr">Koweït</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Open Forum Infectious Diseases</title><idno type="eISSN">2328-8957</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><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></div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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