Figure 2

MyD88, Trif and Caspase-1 are involved in MVAΔIL-1βR-mediated IL-1β induction in murine antigen presenting cells. Bone marrow-derived macrophages (BMDM) (A - C) or myeloid dendritic cells (mDC) (D - F) were generated from C57BL/6 mice (A, D), MyD88^−/−Trif^−/− mice (B, E), or Caspase-1^−/− mice (C, F). Cells were differentiated in vitro from a pool of isolated bone marrow cells derived from two mice per mouse strain. Cells were infected with MVAwt, MVAΔIL-1βR, or MVA-IL-1βRrev at five MOI, mock treated with medium, or incubated in the presence of 1 μg/ml LPS. Infections with each virus were performed once (B, and E) or in duplicate (A, C, D, and F). At 0, 4, 8 and 24 h p.i. cell free supernatants were subjected to IL-1β ELISA. Since vIL-1βR interferes with IL-1β binding by one ELISA antibody, only unbound, free cytokine is measured in this assay. Levels of free IL-1β are shown for each virus from the shortest to the longest time point (left to right). The mean of infection experiments with standard deviations are shown for A, C, D, and F.