Plasmacytoid dendritic cell sensing of African swine fever virus–infected macrophages results in STING-dependent robust interferon-α production

Abstract While several African swine fever virus (ASFV)–encoded proteins potently interfere with the cGAS-STING (cyclic GMP-AMP synthetase–stimulator of interferon genes) pathway at different levels to suppress interferon (IFN) type I production in infected macrophages, systemic IFN-α is induced during the early stages of AFSV infection in pigs. The present study elucidates a mechanism by which such responses can be triggered, at least in vitro. We demonstrate that infection of monocyte-derived macrophages (MDMs) by ASFV genotype 2 strains is highly efficient but immunologically silent with respect to IFN type I, IFN-stimulated gene induction, and tumor necrosis factor production. Additionally, ASFV does not directly activate plasmacytoid dendritic cells (pDCs). However, coculturing pDCs with ASFV-infected MDMs results in a strong pDC response characterized by high levels of IFN-α and tumor necrosis factor. IFN type I, in turn, promoted interleukin-1 receptor antagonist production by macrophages. Similar to the sensing of infected cells by other viruses, pDC activation required integrin-mediated cognate interactions with ASFV-infected MDMs to form an interferogenic synapse. Inhibitor studies indicated that the activation of pDCs requires the STING pathway and the formation of gap junctions. While IL-4–polarized macrophages showed increased susceptibility, IFN-γ–polarized ASFV-infected macrophages induced higher pDC activation. Pretreatment of pDCs with IFN-β and IFN-γ also enhanced IFN-α production in response to ASFV-infected macrophages, highlighting the influence of the immunological microenvironment. These findings suggest that the IFN-α detected during ASFV infection in pigs may be a result of pDC sensing ASFV-infected macrophages.