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Volume 6
Journal of Infectious Diseases and Therapy
ISSN: 2332-0877
Infection Congress 2018
March 01-02, 2018
March 01-02, 2018 Berlin, Germany
5
th
International Congress on
INFECTIOUS DISEASES
J Infect Dis Ther 2018, Volume 6
DOI: 10.4172/2332-0877-C1-039
PCRarrayprofiling of antiviral genes inhuman embryonic kidney cells expressing
Human coronavirus
OC43
structural and accessory proteins
Meshal Samir Beidas
and
Chehadeh W
Kuwait University, Kuwait
Background & Aim:
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 Betacoronavirus 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 defenses 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.
Methods:
HCoV-OC43membrane (M), nucleocapsid (N), ns5a andns2amRNAwere amplified and cloned into the pAcGFP1-N
expression vector (Clontech), followed by transfection in HEK-293 cells. Expressions 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 normalization methods.
Results:
Around 50-60 genes were down-regulated 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 down-regulated genes were those coding for interferons (IFNs) mitogen-
activated protein kinases (MAPKs), pro-apoptotic and pyroptotic proteins (caspases, cathepsins, and tumor necrosis factor),
pro-inflammatory cytokines (interleukins), pattern recognition receptors (PRRs; toll-like receptors and NOD-like receptors)
and their signaling transduction proteins (TICAM1, MAVS).
Conclusion:
This study shows for the first time that similarly to SARS-CoV and MERS-CoV, HCoV-OC43 has the ability to
down-regulate the transcription of genes critical for the activation of different antiviral signaling pathways
.
meshrock2000@hotmail.com