Modified Vaccinia Virus Ankara Triggers Type I IFN Production in Murine Conventional Dendritic Cells via a cGAS/STING-Mediated Cytosolic DNA-Sensing Pathway
Figure 5
cGAS is the critical cytosolic DNA sensor for MVA infection of cDCs.
GM-CSF-BMDCs were generated from cGAS−/− mice and its age-matched WT controls. (A) Cells (1×106) were infected with MVA at a MOI of 10. Supernatants were collected 22 h later. The concentrations of IFN-α and IFN-β were determined by ELISA. Data are means ± SEM (n = 3). A representative experiment is shown, repeated twice (***, p<0.001). (B) Cells (1×106) were infected with MVA at a MOI of 10. Cells were collected at 6 h post infection. Real-time PCR analysis of IFNA4 and IFNB mRNAs were performed. Data are means ± SEM (n = 3). A representative experiment is shown, repeated twice (***, p<0.001). (C) Western blot analysis of cGAS+/+ and cGAS−/− cDCs infected with MVA at a MOI of 10, or mock infected. Whole-cell lysates were prepared. Equal amount of proteins were subjected to SDS-PAGE and immunoblotting with anti-phospho-TBK1, anti-TBK1, anti-phosphoserine-396 of IRF3, and anti-IRF3. GAPDH was used as a loading control. “hpi”, hours post infection. “M”, mock infection control.