Cytokine secretion and activation of CD11c⁺ splenocytes, IPCs, CD11b⁺ DC, CD11b^– DCs, and bone marrow–derived IPCs from wild-type and MyD88–/– mice in response to HSV-1 in vitro. (A) CD11c⁺ splenocytes were isolated from wild-type (▪) and MyD88–/– (▨) mice and cultured for 36 hours (2 × 10⁶ cells/mL) with KOS HSV-1 at the indicated MOI or 3 μg/mL CpG ODN 2216. (B) IPC (top row), CD11b⁺ DCs (middle row), and CD11b^– DC (bottom row) were sorted from wild-type and MyD88–/– CD11c⁺ splenocytes and were cultured with 2 MOI HSV-1 or 3 μg/mL CpG ODN 2216 for 36 hours (5 × 10⁴ cells/150 μL/well). (C) Bone marrow–derived wild-type and MyD88–/– IPCs were incubated with HSV-1 (0.04-5 MOI) or CpG ODN 2216 (3 μg/mL) for 36 hours at 10⁶ cells/mL. IFN-α and IL-12 were measured in the supernatants using ELISA. CD86 expression was measured as a marker of activation by flow cytometry and is expressed as median fluorescence intensity. Dashed line indicates the median value of the isotype control sample.