Monocyte-derived macrophage stimulation

Monocyte-derived macrophages (MDMs) were treated with muramyl dipeptide (MDP; Bachem, Torrance, CA), lipid A (Peptides International, Louisville, KY), Pam3Cys (EMD Millipore, Billerica, MA), polyI:C, flagellin, CL097, CpG (InvivoGen, San Diego, CA), IL23 (PeproTech), 10:1 multiplicity of infection Salmonella enterica serovar Typhimurium or adherent invasive Escherichia coli (AIEC) strain LF82. In some cases cells were incubated with neutralising anti-IL23p19 antibody (eBioscience, San Diego, CA) for 1 hour before the treatments. Supernatants were assayed for tumour necrosis factor (TNF), IL6, IL8, IL10 (BD Biosciences, San Jose, CA), IL1β or IL23 (coating antibody to IL23p19, clone eBio473P19, detection antibody to IL12/ IL23p40, clone C8.6) (eBioscience) by ELISA.