Innate immune response of human Plasmacytoid dendritic cells to poxvirus infection is subverted by vaccinia E3 via its Z-DNA/RNA binding domain

Hua Cao, Peihong Dai, Weiyi Wang, Hao Li, Jianda Yuan, Fangjin Wang, Chee Mun Fang, Paula M. Pitha, Jia Liu, Richard C. Condit, Douglas McFadden, Taha Merghoub, Alan N. Houghton, James W. Young, Stewart Shuman, Liang Deng

Research output: Contribution to journalArticle

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Abstract

Plasmacytoid dendritic cells (pDCs) play important roles in antiviral innate immunity by producing type I interferon (IFN). In this study, we assess the immune responses of primary human pDCs to two poxviruses, vaccinia and myxoma virus. Vaccinia, an orthopoxvirus, was used for immunization against smallpox, a contagious human disease with high mortality. Myxoma virus, a Leporipoxvirus, causes lethal disease in rabbits, but is non-pathogenic in humans. We report that myxoma virus infection of human pDCs induces IFN-α and TNF production, whereas vaccinia infection does not. Co-infection of pDCs with myxoma virus plus vaccinia blocks myxoma induction effects. We find that heat-inactivated vaccinia (Heat-VAC; by incubating the virus at 55°C for 1 h) gains the ability to induce IFN-α and TNF in primary human pDCs. Induction of IFN-α in pDCs by myxoma virus or Heat-VAC is blocked by chloroquine, which inhibits endosomal acidification required for TLR7/9 signaling, and by inhibitors of cellular kinases PI3K and Akt. Using purified pDCs from genetic knockout mice, we demonstrate that Heat-VAC-induced type I IFN production in pDCs requires the endosomal RNA sensor TLR7 and its adaptor MyD88, transcription factor IRF7 and the type I IFN feedback loop mediated by IFNAR1. These results indicate that (i) vaccinia virus, but not myxoma virus, expresses inhibitor(s) of the poxvirus sensing pathway(s) in pDCs; and (ii) Heat-VAC infection fails to produce inhibitor(s) but rather produces novel activator(s), likely viral RNA transcripts that are sensed by the TLR7/MyD88 pathway. Using vaccinia gene deletion mutants, we show that the Z-DNA/RNA binding domain at the N-terminus of the vaccinia immunomodulatory E3 protein is an antagonist of the innate immune response of human pDCs to poxvirus infection and TLR agonists. The myxoma virus ortholog of vaccinia E3 (M029) lacks the N-terminal Z-DNA/RNA binding domain, which might contribute to the immunostimulating properties of myxoma virus.

Original languageEnglish (US)
Article numbere36823
JournalPLoS One
Volume7
Issue number5
DOIs
StatePublished - May 14 2012
Externally publishedYes

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Poxviridae Infections
Z-DNA
Poxviridae
Z-Form DNA
Myxoma virus
Vaccinia
Vaccinium
dendritic cells
Viruses
Innate Immunity
Dendritic Cells
RNA
interferons
infection
Hot Temperature
Interferon Type I
heat
Interferons
Vaccinia virus
Leporipoxvirus

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Innate immune response of human Plasmacytoid dendritic cells to poxvirus infection is subverted by vaccinia E3 via its Z-DNA/RNA binding domain. / Cao, Hua; Dai, Peihong; Wang, Weiyi; Li, Hao; Yuan, Jianda; Wang, Fangjin; Fang, Chee Mun; Pitha, Paula M.; Liu, Jia; Condit, Richard C.; McFadden, Douglas; Merghoub, Taha; Houghton, Alan N.; Young, James W.; Shuman, Stewart; Deng, Liang.

In: PLoS One, Vol. 7, No. 5, e36823, 14.05.2012.

Research output: Contribution to journalArticle

Cao, H, Dai, P, Wang, W, Li, H, Yuan, J, Wang, F, Fang, CM, Pitha, PM, Liu, J, Condit, RC, McFadden, D, Merghoub, T, Houghton, AN, Young, JW, Shuman, S & Deng, L 2012, 'Innate immune response of human Plasmacytoid dendritic cells to poxvirus infection is subverted by vaccinia E3 via its Z-DNA/RNA binding domain', PLoS One, vol. 7, no. 5, e36823. https://doi.org/10.1371/journal.pone.0036823
Cao, Hua ; Dai, Peihong ; Wang, Weiyi ; Li, Hao ; Yuan, Jianda ; Wang, Fangjin ; Fang, Chee Mun ; Pitha, Paula M. ; Liu, Jia ; Condit, Richard C. ; McFadden, Douglas ; Merghoub, Taha ; Houghton, Alan N. ; Young, James W. ; Shuman, Stewart ; Deng, Liang. / Innate immune response of human Plasmacytoid dendritic cells to poxvirus infection is subverted by vaccinia E3 via its Z-DNA/RNA binding domain. In: PLoS One. 2012 ; Vol. 7, No. 5.
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N2 - Plasmacytoid dendritic cells (pDCs) play important roles in antiviral innate immunity by producing type I interferon (IFN). In this study, we assess the immune responses of primary human pDCs to two poxviruses, vaccinia and myxoma virus. Vaccinia, an orthopoxvirus, was used for immunization against smallpox, a contagious human disease with high mortality. Myxoma virus, a Leporipoxvirus, causes lethal disease in rabbits, but is non-pathogenic in humans. We report that myxoma virus infection of human pDCs induces IFN-α and TNF production, whereas vaccinia infection does not. Co-infection of pDCs with myxoma virus plus vaccinia blocks myxoma induction effects. We find that heat-inactivated vaccinia (Heat-VAC; by incubating the virus at 55°C for 1 h) gains the ability to induce IFN-α and TNF in primary human pDCs. Induction of IFN-α in pDCs by myxoma virus or Heat-VAC is blocked by chloroquine, which inhibits endosomal acidification required for TLR7/9 signaling, and by inhibitors of cellular kinases PI3K and Akt. Using purified pDCs from genetic knockout mice, we demonstrate that Heat-VAC-induced type I IFN production in pDCs requires the endosomal RNA sensor TLR7 and its adaptor MyD88, transcription factor IRF7 and the type I IFN feedback loop mediated by IFNAR1. These results indicate that (i) vaccinia virus, but not myxoma virus, expresses inhibitor(s) of the poxvirus sensing pathway(s) in pDCs; and (ii) Heat-VAC infection fails to produce inhibitor(s) but rather produces novel activator(s), likely viral RNA transcripts that are sensed by the TLR7/MyD88 pathway. Using vaccinia gene deletion mutants, we show that the Z-DNA/RNA binding domain at the N-terminus of the vaccinia immunomodulatory E3 protein is an antagonist of the innate immune response of human pDCs to poxvirus infection and TLR agonists. The myxoma virus ortholog of vaccinia E3 (M029) lacks the N-terminal Z-DNA/RNA binding domain, which might contribute to the immunostimulating properties of myxoma virus.

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