Fig. 2.
Response of HPMC to bacterial TLR agonists. (A) HPMC were stimulated with serial dilutions of SES (1/200-Neat) Pam3Cys (0.1–20 μg/mL) or LPS (2–500 ng/mL). After 24-h incubation CXCL8, IL-6 or CCL2 levels were quantified in cell-free supernatants using ELISA. Data shown are a result representative of at least three experiments performed using primary cell lines from different donors (*P < 0.05; **P < 0.01 between unstimulated and stimulated cells with the indicated ligands). (B) HPMC were stimulated with high doses of LPS (1–5 μg/mL). After 24-h incubation, CXCL8 and CCL5 levels were quantified in cell-free supernatants using ELISA (***P < 0.001). (C) HPMC were stimulated for 24 h with increasing doses of Pam2Cys (MALP-2 analogue) or neat SES (with 0.1% FCS) and CXCL8 levels were quantified by ELISA in cell-free supernatants. Data presented are results representative of four independent experiments (**P < 0.01; P < 0.001 between unstimulated and stimulated cells with Pam2Cys. (D) HPMC were stimulated for 24 h with increasing doses of flagellin and CXCL8 levels were quantified by ELISA in cell-free supernatants. Data presented are results representative of two independent experiments (*P < 0.05 between unstimulated and stimulated cells with flagellin).