FIG 5.

Chemokine production by M1 macrophages in response to Porphyromonas gulae whole cells. WT iMACs, TLR2^−/− iMACs, and TLR4^−/− iMACs (10⁵ cells) were incubated overnight with IFN-γ to prime the M1 macrophage phenotype. The M1-primed macrophages were then incubated with viable P. gulae (MOI of 180:1) for 2 h and washed with DMEM containing antibiotics. E. coli LPS and Pam3CSK4 were used as TLR4 and TLR2 control ligands, respectively. The negative control was IFN-γ-primed macrophages that were not incubated with TLR ligands or P. gulae cells. After 24 h, the levels of chemokines in the assay supernatants were measured using a 23-plex Bioplex assay. The assay was repeated twice on each of three biological replicates (n = 6). (a) Eotaxin; (b) G-CSF; (c) KC; (d) MCP-1; (e) MIP-1α; (f) RANTES. *, P < 0.05 compared to the corresponding negative control.