Role of gut microbiota in modulation of dendritic cell mediated immune tolerance with age

With our experiments we have shown LP(Lactobacillus plantarum) mediated modification of immunogenic bone marrow derived dendritic cells (BMDCs) to acquire tolerogenic phenotype, to unravel the molecular mechanism of LP mediated immunomodulation we performed global transcriptomic Gut of old mice was reconstituted with LP, and RNA was extracted from LPS stimulated BMDCs from young, young treated with antibiotic cocktail, old and old-LP groups for gene array profiling by Agilent GeneChips. TRIzol reagent was used to isolate total RNA from DCs after LPS stimulation (1μg/ml) for 24 hours according to the manufacturer's protocol (Invitrogen, CA, USA). Later, DNase1 enzyme was added to digest and remove genomic DNA. Concentrations of RNA were measured with the NanoDrop ND-100 Spectrophotometer