PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins
Identifieur interne : 000636 ( Pmc/Curation ); précédent : 000635; suivant : 000637PCR 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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<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>
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<author><name sortKey="Chehadeh, Wassim" sort="Chehadeh, Wassim" uniqKey="Chehadeh W" first="Wassim" last="Chehadeh">Wassim Chehadeh</name>
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,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
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,<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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<series><title level="j">Open Forum Infectious Diseases</title>
<idno type="eISSN">2328-8957</idno>
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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>
</div>
</front>
</TEI>
<pmc article-type="abstract"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Open Forum Infect Dis</journal-id>
<journal-id journal-id-type="iso-abbrev">Open Forum Infect Dis</journal-id>
<journal-id journal-id-type="publisher-id">ofid</journal-id>
<journal-title-group><journal-title>Open Forum Infectious Diseases</journal-title>
</journal-title-group>
<issn pub-type="epub">2328-8957</issn>
<publisher><publisher-name>Oxford University Press</publisher-name>
<publisher-loc>US</publisher-loc>
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<article-id pub-id-type="doi">10.1093/ofid/ofx163.728</article-id>
<article-id pub-id-type="publisher-id">ofx163.728</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Abstracts</subject>
<subj-group subj-group-type="category-toc-heading"><subject>Poster Abstract</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group><article-title>PCR Array Profiling of Antiviral Genes in Human Embryonic Kidney Cells Expressing Human Coronavirus OC43 Structural and Accessory Proteins</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Beidas</surname>
<given-names>Meshal</given-names>
</name>
<degrees>MSc</degrees>
<xref ref-type="aff" rid="AF0001">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chehadeh</surname>
<given-names>Wassim</given-names>
</name>
<degrees>PhD</degrees>
<xref ref-type="aff" rid="AF0001">1</xref>
</contrib>
</contrib-group>
<aff id="AF0001"><label>1</label>
<institution>Microbiology, Kuwait University</institution>
,<addr-line>Kuwait</addr-line>
,<country>Kuwait</country>
</aff>
<author-notes><fn id="fn-0001"><p><bold>Session:</bold>
139. Adult Viral Infection</p>
<p><italic>Friday, October 6, 2017: 12:30 PM</italic>
</p>
</fn>
</author-notes>
<pub-date pub-type="collection"><season>Fall</season>
<year>2017</year>
</pub-date>
<pub-date pub-type="epub" iso-8601-date="2017-10-04"><day>04</day>
<month>10</month>
<year>2017</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>04</day>
<month>10</month>
<year>2017</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on the . </pmc-comment>
<volume>4</volume>
<issue>Suppl 1</issue>
<issue-title>ID Week 2017 Abstracts</issue-title>
<fpage>S312</fpage>
<lpage>S312</lpage>
<permissions><copyright-statement>© The Author 2017. Published by Oxford University Press on behalf of Infectious Diseases Society of America.</copyright-statement>
<copyright-year>2017</copyright-year>
<license license-type="cc-by-nc-nd" xlink:href="http://creativecommons.org/licenses/by-nc-nd/4.0"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc-nd/4.0/">http://creativecommons.org/licenses/by-nc-nd/4.0/</ext-link>
), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com</license-p>
</license>
</permissions>
<self-uri xlink:href="ofx163.728.pdf"></self-uri>
<abstract><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>
</abstract>
<counts><page-count count="1"></page-count>
</counts>
</article-meta>
</front>
</pmc>
</record>
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