Gene expression analysis of PBMC's from IBD subjects stimulated by combinations of LPS, MDP, anti-CD3/anti-CD28 antibodies, and anti-IL10R antibodies.

We systematically investigated the transcriptomic response pattern of PBMC from 41 patients with IBD to characterize relevant stimuli that target different aspects of innate and adaptive immune cell responses. Lipopolysaccharide(LPS), muramyl-dipeptide (L18MDP), T cell receptor and co-stimulation (αCD3/αCD28 coated beads), and IL-10 signalling blockade were used to sitmulate subject PBMC's alone or in combination based on the concept that innate pathogen recognition receptor responses (in particular NOD2 and TLR4), T cell responses, and IL-10 signalling defects are implicated by multiple genetic IBD susceptibility loci and Mendelian forms of IBD. PBMC's isolated from IBD patients were subjected to microarray gene expression analysis after 16 hours of stimulation (by LPS, L18MDP and anti-CD3/anti-CD28 coated beads) in the presence or absence of IL-10 receptor-blocking antibodies.