Production of type I IFNs, pro-inflammatory cytokines and β-chemokines after TLR stimulation in macrophages.

<p>MDMs were cultured in quadruplicate for 16 h unstimulated or with the ligands for TLR2, TLR3, TLR4, TLR7 or TLR9, and then supernatants were collected, clarified, aliquoted and stored at −80°C until use. Commercial enzyme-linked immunoassays were used for quantitation of type I IFNs IFNα (PBL InterferonSource and Thermo Scientific) (A) and IFNβ (Thermo Scientific) (B), pro-inflammatory cytokines TNF-α (C) and IL-6 (D) (both from eBioscience), and β-chemokines MIP-1α (CCL3) (E) (SABiosciences), MIP-1β (CCL4) (F) (Mabtech) and RANTES (CCL5) (G) (Peprotech). All samples were analyzed in duplicate. Multiple donors are shown for type I IFNs and one representative donor is shown for pro-inflammatory cytokines and β-chemokines. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002937#s3" target="_blank">Results</a> are shown as mean ± SD of duplicate cultures. Dashed lines indicate the limit of detection of the assays for quantitation of type I IFNs.