Gene expression in the small intestine of mice colonized with Pseudomonas aeruginosa

Celiac disease (CeD) is a food sensitivity characterized by a breakdown of oral tolerance to gluten proteins in genetically predisposed individuals, although the underlying mechanisms are incompletely understood. To evaluate a functional role of bacterial proteases in CeD, we colonized germ-free or clean SPF mice with P.aeruginosa PA14 or P.aeruginosa PA14 disrupted in lasB gene (elastase). We show gluten-independent, PAR-2 mediated upregulation of inflammatory pathways by LasB in C57BL/6 mice without villus blunting. In mice expressing CeD risk genes, P. aeruginosa elastase synergized with gluten to induce more severe inflammation that was associated with moderate villus blunting. Determination of inflammatory genes in tissue and intraepithelial lymphocytes of mice. For RNA extraction, RNeasy Mini Kit (Qiagen) was used. Gene Expression CodeSet for mouse inflammation genes (Mouse Inflammation panel, 254 genes) were run according to the manufacturer’s instructions (NanoString Technologies Inc.). The results obtained were analyzed with nSolver 2.5 (NanoString Technologies).