Description

Isolating bacteriophages from a Crohn’s Disease patient: Inflammatory bowel disease (IBD) encompasses two conditions, Crohn's disease (CD) and ulcerative colitis (UC), and is characterized by inflammation of the gastrointestinal tract. These diseases are correlated with a decrease in the microbial diversity in the gut. While much is known about the role of the microbiome in IBD, the role of the virome–specifically how bacteriophages modulate the gut microbiome–is poorly understood. We hypothesize that a bloom of lytic or temperate phages in the gut plays a role in modulation of the microbiome and the consequent inflammatory response of the human host, and therefore impacts disease state and progression. To address this hypothesis, we have isolated bacteria from patients with IBD and house-hold controls (HHC) in anaerobic conditions. We then use the bacterial isolates to identify and characterize bacteriophages found in the gut from either diseased or healthy individuals. The isolation and characterization of anaerobic bacteriophages remains difficult, and many fewer phages are known for anaerobic than aerobic bacteria. In this presentation, we will show our approach for isolating bacteria and creating phage lysates from the stools of IBD patients and matched healthy individuals using both classic and enrichment strategies. We will also show that bacteriophages isolated from a Crohn’s disease patient are able to plaque on bacterial isolates from the same individual. Our goal is to identify which phages are positively or negatively associated with disease, and the mechanism by which they promote the disease or protect from it. Funding: NIDDK RC2DK116713 Computational and Experimental Resources for Virome Analysis in Inflammatory Bowel Disease