Middle East respiratory syndrome coronavirus shows poor replication but significant induction of antiviral responses in human monocyte-derived macrophages and dendritic cells
In this study we assessed the ability of Middle East respiratory syndrome coronavirus (MERS-CoV) to replicate and induce innate immunity in human monocyte-derived macrophages and dendritic cells (MDDCs), and compared it with severe acute respiratory syndrome coronavirus (SARS-CoV). Assessments of viral protein and RNA levels in infected cells showed that both viruses were impaired in their ability to replicate in these cells. Some induction of IFN-λ1, CXCL10 and MxA mRNAs in both macrophages and MDDCs was seen in response to MERS-CoV infection, but almost no such induction was observed in response to SARS-CoV infection. ELISA and Western blot assays showed clear production of CXCL10 and MxA in MERS-CoV-infected macrophages and MDDCs. Our data suggest that SARS-CoV and MERS-CoV replicate poorly in human macrophages and MDDCs, but MERS-CoV is nonetheless capable of inducing a readily detectable host innate immune response. Our results highlight a clear difference between the viruses in activating host innate immune responses in macrophages and MDDCs, which may contribute to the pathogenesis of infection.
AdneyD. R.,
van DoremalenN.,
BrownV. R.,
BushmakerT.,
ScottD.,
de WitE.,
BowenR. A.,
MunsterV. J.2014; Replication and shedding of MERS-CoV in upper respiratory tract of inoculated dromedary camels. Emerg Infect Dis 20:1999–2005 [View Article][PubMed]
AzharE. I.,
El-KafrawyS. A.,
FarrajS. A.,
HassanA. M.,
Al-SaeedM. S.,
HashemA. M.,
MadaniT. A.2014; Evidence for camel-to-human transmission of MERS coronavirus. N Engl J Med 370:2499–2505 [View Article][PubMed]
BoschB. J.,
van der ZeeR.,
de HaanC. A.,
RottierP. J.2003; The coronavirus spike protein is a class I virus fusion protein: structural and functional characterization of the fusion core complex. J Virol 77:8801–8811 [View Article][PubMed]
BurkardC.,
VerheijeM. H.,
WichtO.,
van KasterenS. I.,
van KuppeveldF. J.,
HaagmansB. L.,
PelkmansL.,
RottierP. J.,
BoschB. J.,
de HaanC. A.2014; Coronavirus cell entry occurs through the endo-/lysosomal pathway in a proteolysis-dependent manner. PLoS Pathog 10:e1004502 [View Article][PubMed]
ChanJ. F.,
ChanK. H.,
ChoiG. K.,
ToK. K.,
TseH.,
CaiJ. P.,
YeungM. L.,
ChengV. C.,
ChenH.,
other authors.
2013; Differential cell line susceptibility to the emerging novel human betacoronavirus 2c EMC/2012: implications for disease pathogenesis and clinical manifestation. J Infect Dis 207:1743–1752 [View Article][PubMed]
ChengV. C. C.,
LauS. K. P.,
WooP. C. Y.,
YuenK. Y.2007; Severe acute respiratory syndrome coronavirus as an agent of emerging and reemerging infection. Clin Microbiol Rev 20:660–694 [View Article][PubMed]
CormanV. M.,
EckerleI.,
BleickerT.,
ZakiA.,
LandtO.,
Eschbach-BludauM.,
van BoheemenS.,
GopalR.,
BallhauseM.2012; Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction. Euro Surveill 17:3–8[PubMed]
CormanV. M.,
ItheteN. L.,
RichardsL. R.,
SchoemanM. C.,
PreiserW.,
DrostenC.,
DrexlerJ. F.2014; Rooting the phylogenetic tree of Middle East respiratory syndrome coronavirus by characterization of a conspecific virus from an African bat. J Virol 88:11297–11303 [View Article][PubMed]
de GrootR. J.,
BakerS. C.,
BaricR. S.,
BrownC. S.,
DrostenC.,
EnjuanesL.,
FouchierR. A. M.,
GalianoM.,
GorbalenyaA. E.,
other authors.
2013; Middle East respiratory syndrome coronavirus (MERS-CoV): announcement of the Coronavirus Study Group. J Virol 87:7790–7792 [View Article][PubMed]
DeDiegoM. L.,
Nieto-TorresJ. L.,
Jimenez-GuardeñoJ. M.,
Regla-NavaJ. A.,
Castaño-RodriguezC.,
Fernandez-DelgadoR.,
UseraF.,
EnjuanesL.2014; Coronavirus virulence genes with main focus on SARS-CoV envelope gene. Virus Res 194:124–137 [View Article][PubMed]
DrostenC.,
GüntherS.,
PreiserW.,
van der WerfS.,
BrodtH. R.,
BeckerS.,
RabenauH.,
PanningM.,
KolesnikovaL.,
other authors.
2003; Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 348:1967–1976 [View Article][PubMed]
FaureE.,
PoissyJ.,
GoffardA.,
FournierC.,
KipnisE.,
TitecatM.,
BortolottiP.,
MartinezL.,
DubucquoiS.,
other authors.
2014; Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside?. PLoS One 9:e88716 [View Article][PubMed]
FunkC. J.,
WangJ.,
ItoY.,
TravantyE. A.,
VoelkerD. R.,
HolmesK. V.,
MasonR. J.2012; Infection of human alveolar macrophages by human coronavirus strain 229E. J Gen Virol 93:494–503 [View Article][PubMed]
GiererS.,
BertramS.,
KaupF.,
WrenschF.,
HeurichA.,
Krämer-KühlA.,
WelschK.,
WinklerM.,
MeyerB.,
other authors.
2013; The spike protein of the emerging betacoronavirus EMC uses a novel coronavirus receptor for entry, can be activated by TMPRSS2, and is targeted by neutralizing antibodies. J Virol 87:5502–5511 [View Article][PubMed]
HuiD. S.,
MemishZ. A.,
ZumlaA.2014; Severe acute respiratory syndrome vs. the Middle East respiratory syndrome. Curr Opin Pulm Med 20:233–241 [View Article][PubMed]
ItheteN. L.,
StoffbergS.,
CormanV. M.,
CottontailV. M.,
RichardsL. R.,
SchoemanM. C.,
DrostenC.,
DrexlerJ. F.,
PreiserW.2013; Close relative of human Middle East respiratory syndrome coronavirus in bat, South Africa. Emerg Infect Dis 19:1697–1699 [View Article][PubMed]
JossetL.,
MenacheryV. D.,
GralinskiL. E.,
AgnihothramS.,
SovaP.,
CarterV. S.,
YountB. L.,
GrahamR. L.,
BaricR. S.,
KatzeM. G.2013; Cell host response to infection with novel human coronavirus EMC predicts potential antivirals and important differences with SARS coronavirus. MBio 4:e00165–15 [View Article][PubMed]
Kopecky-BrombergS. A.,
Martínez-SobridoL.,
FriemanM.,
BaricR. A.,
PaleseP.2007; Severe acute respiratory syndrome coronavirus open reading frame (ORF) 3b, ORF 6, and nucleocapsid proteins function as interferon antagonists. J Virol 81:548–557 [View Article][PubMed]
KopfM.,
SchneiderC.,
NobsS. P.2015; The development and function of lung-resident macrophages and dendritic cells. Nat Immunol 16:36–44 [View Article][PubMed]
LambeirA. M.,
DurinxC.,
ScharpéS.,
De MeesterI.2003; Dipeptidyl-peptidase IV from bench to bedside: an update on structural properties, functions, and clinical aspects of the enzyme DPP IV. Crit Rev Clin Lab Sci 40:209–294 [View Article][PubMed]
LauS. K.,
LauC. C.,
ChanK. H.,
LiC. P.,
ChenH.,
JinD. Y.,
ChanJ. F.,
WooP. C.,
YuenK. Y.2013; Delayed induction of proinflammatory cytokines and suppression of innate antiviral response by the novel Middle East respiratory syndrome coronavirus: implications for pathogenesis and treatment. J Gen Virol 94:2679–2690 [View Article][PubMed]
LehtonenA.,
AhlforsH.,
VeckmanV.,
MiettinenM.,
LahesmaaR.,
JulkunenI.2007; Gene expression profiling during differentiation of human monocytes to macrophages or dendritic cells. J Leukoc Biol 82:710–720 [View Article][PubMed]
LiW.,
MooreM. J.,
VasilievaN.,
SuiJ.,
WongS. K.,
BerneM. A.,
SomasundaranM.,
SullivanJ. L.,
LuzuriagaK.,
other authors.
2003; Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426:450–454 [View Article][PubMed]
MatthewsK. L.,
ColemanC. M.,
van der MeerY.,
SnijderE. J.,
FriemanM. B.2014; The ORF4b-encoded accessory proteins of Middle East respiratory syndrome coronavirus and two related bat coronaviruses localize to the nucleus and inhibit innate immune signalling. J Gen Virol 95:874–882 [View Article][PubMed]
MeyerB.,
MüllerM. A.,
CormanV. M.,
ReuskenC. B.,
RitzD.,
GodekeG. J.,
LattweinE.,
KalliesS.,
SiemensA.,
other authors.
2014; Antibodies against MERS coronavirus in dromedary camels, United Arab Emirates, 2003 and 2013. Emerg Infect Dis 20:552–559 [View Article][PubMed]
MilletJ. K.,
WhittakerG. R.2014; Host cell entry of Middle East respiratory syndrome coronavirus after two-step, furin-mediated activation of the spike protein. Proc Natl Acad Sci U S A 111:15214–15219 [View Article][PubMed]
MinakshiR.,
PadhanK.,
RaniM.,
KhanN.,
AhmadF.,
JameelS.2009; The SARS Coronavirus 3a protein causes endoplasmic reticulum stress and induces ligand-independent downregulation of the type 1 interferon receptor. PLoS One 4:e8342 [View Article][PubMed]
NiemeyerD.,
ZillingerT.,
MuthD.,
ZieleckiF.,
HorvathG.,
SulimanT.,
BarchetW.,
WeberF.,
DrostenC.,
MüllerM. A.2013; Middle East respiratory syndrome coronavirus accessory protein 4a is a type I interferon antagonist. J Virol 87:12489–12495 [View Article][PubMed]
OsterlundP.,
VeckmanV.,
SirénJ.,
KlucherK. M.,
HiscottJ.,
MatikainenS.,
JulkunenI.2005; Gene expression and antiviral activity of alpha/beta interferons and interleukin-29 in virus-infected human myeloid dendritic cells. J Virol 79:9608–9617 [View Article][PubMed]
OsterlundP.,
PirhonenJ.,
IkonenN.,
RönkköE.,
StrengellM.,
MäkeläS. M.,
BromanM.,
HammingO. J.,
HartmannR.,
other authors.
2010; Pandemic H1N1 2009 influenza A virus induces weak cytokine responses in human macrophages and dendritic cells and is highly sensitive to the antiviral actions of interferons. J Virol 84:1414–1422 [View Article][PubMed]
PirhonenJ.,
SarenevaT.,
KurimotoM.,
JulkunenI.,
MatikainenS.1999; Virus infection activates IL-1 beta and IL-18 production in human macrophages by a caspase-1-dependent pathway. J Immunol 162:7322–7329[PubMed]
RajV. S.,
MouH.,
SmitsS. L.,
DekkersD. H. W.,
MüllerM. A.,
DijkmanR.,
MuthD.,
DemmersJ. A. A.,
ZakiA.,
other authors.
2013; Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature 495:251–254 [View Article][PubMed]
RajV. S.,
FaragE. A.,
ReuskenC. B.,
LamersM. M.,
PasS. D.,
VoermansJ.,
SmitsS. L.,
OsterhausA. D.,
Al-MawlawiN.,
other authors.
2014; Isolation of MERS coronavirus from a dromedary camel, Qatar, 2014. Emerg Infect Dis 20:1339–1342 [View Article][PubMed]
ScheupleinV. A.,
SeifriedJ.,
MalczykA. H.,
MillerL.,
HöckerL.,
Vergara-AlertJ.,
DolnikO.,
ZieleckiF.,
BeckerB.,
other authors.
2015; High secretion of interferons by human plasmacytoid dendritic cells upon recognition of Middle East respiratory syndrome coronavirus. J Virol 89:3859–3869 [View Article][PubMed]
ShortK. R.,
BrooksA. G.,
ReadingP. C.,
LondriganS. L.2012; The fate of influenza A virus after infection of human macrophages and dendritic cells. J Gen Virol 93:2315–2325 [View Article][PubMed]
SiuK. L.,
KokK. H.,
NgM. H.,
PoonV. K.,
YuenK. Y.,
ZhengB. J.,
JinD. Y.2009; Severe acute respiratory syndrome coronavirus M protein inhibits type I interferon production by impeding the formation of TRAF3·TANK·TBK1/IKKepsilon complex. J Biol Chem 284:16202–16209 [View Article][PubMed]
SiuK. L.,
YeungM. L.,
KokK. H.,
YuenK. S.,
KewC.,
LuiP. Y.,
ChanC. P.,
TseH.,
WooP. C.,
other authors.
2014; Middle East respiratory syndrome coronavirus 4a protein is a double-stranded RNA-binding protein that suppresses PACT-induced activation of RIG-I and MDA5 in the innate antiviral response. J Virol 88:4866–4876 [View Article][PubMed]
SpiegelM.,
PichlmairA.,
Martínez-SobridoL.,
CrosJ.,
García-SastreA.,
HallerO.,
WeberF.2005; Inhibition of beta interferon induction by severe acute respiratory syndrome coronavirus suggests a two-step model for activation of interferon regulatory factor 3. J Virol 79:2079–2086 [View Article][PubMed]
WangQ.,
QiJ.,
YuanY.,
XuanY.,
HanP.,
WanY.,
JiW.,
LiY.,
WuY.,
other authors.
2014; Bat origins of MERS-CoV supported by bat coronavirus HKU4 usage of human receptor CD26. Cell Host Microbe 16:328–337 [View Article][PubMed]
YangY.,
ZhangL.,
GengH.,
DengY.,
HuangB.,
GuoY.,
ZhaoZ.,
TanW.2013; The structural and accessory proteins M, ORF 4a, ORF 4b, and ORF 5 of Middle East respiratory syndrome coronavirus (MERS-CoV) are potent interferon antagonists. Protein Cell 4:951–961 [View Article][PubMed]
ZakiA. M.,
van BoheemenS.,
BestebroerT. M.,
OsterhausA.D.M.E.,
FouchierR. A. M.2012; Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med 367:1814–1820 [View Article][PubMed]
ZhouJ.,
ChuH.,
LiC.,
WongB. H.,
ChengZ. S.,
PoonV. K.,
SunT.,
LauC. C.,
WongK. K.,
other authors.
2014; Active replication of Middle East respiratory syndrome coronavirus and aberrant induction of inflammatory cytokines and chemokines in human macrophages: implications for pathogenesis. J Infect Dis 209:1331–1342[PubMed][CrossRef]
ZieleckiF.,
WeberM.,
EickmannM.,
SpiegelbergL.,
ZakiA. M.,
MatrosovichM.,
BeckerS.,
WeberF.2013; Human cell tropism and innate immune system interactions of human respiratory coronavirus EMC compared to those of severe acute respiratory syndrome coronavirus. J Virol 87:5300–5304 [View Article][PubMed]
Middle East respiratory syndrome coronavirus shows poor replication but significant induction of antiviral responses in human monocyte-derived macrophages and dendritic cells