CD200R1 licenses pro-inflammatory signaling in peritoneal macrophages.

<p>(A) CD200R1^+/+ (WT) or CD200R1^−/− (KO) elicited peritoneal macrophages were stimulated with either HSV-1 (MOI 10∶1, left column), Pam₂CSK₄ (100 ng/ml, center column), LPS (100 ng/ml, right column), or medium for 24, 48, or 72 h. IL-6 (upper row, pg/ml), CCL5/Rantes (middle row, pg/ml), and CCL2/MCP1 (lower row, pg/ml) levels were measured by ELISA. Data shown are mean and SD of representative experiment (total of 3 experiments, n = 4, WT and KO). An unpaired, two tailed Student’s t-test was used to determine statistical significance of independent experiments, p values; * p<0.05, † p<0.01. (B) Supernatant IL-1β levels from WT and KO elicited peritoneal macrophages (left panels) or bone marrow macrophages (right panels) 16 h after addition of HSV-1 (MOI 10∶1) or after a 3 h stimulation with LPS (100 ng/ml) followed by ATP (1 mM) for 1 h. ELISA results are representative of 3 experiments. (C) WT and KO elicited peritoneal macrophages were cultured for 20 h with HSV-1 (MOI 10∶1). Media (left lanes) and cells (right lanes) were analyzed by SDS-PAGE followed by Western blot for cleavage of pro-IL-1β to IL-1β. Data representative of 3 experiments (n = 4, WT; n = 5, KO).