ORIGINAL ARTICLE Ecobiotherapy Rich in Firmicutes Decreases Susceptibility to Colitis in a Humanized Gnotobiotic Mouse Model Jane M. Natividad, PhD,* Maria I. Pinto-Sanchez, MD,* Heather J. Galipeau, PhD,* Jennifer Jury, MSc,* Manel Jordana, MD, PhD, Walter Reinisch, MD,* Stephen M. Collins, MD,* Premsyl Bercik, MD,* Michael G. Surette, PhD,* Emma Allen-Vercoe, PhD, and Elena F. Verdu, MD, PhD* Background: Alterations in the intestinal microbiota, characterized by depletion of anti-inammatory bacteria, such as Firmicutes, in patients with ulcerative colitis (UC) have prompted interest in microbiota-modulating strategies for this condition. The aim of this study was to evaluate the role of fecal and synthetic human microbial ecosystems, low or enriched in Firmicutes, on colitis susceptibility and host immune responses. Methods: The microbiota of selected healthy and UC human donors was characterized by culture method and 16S rRNA-based sequencing. Germ-free mice were colonized with fecal or a synthetic ecosystem enriched (healthy donors) or low (UC donors) in Firmicutes. Experimental colitis was induced using dextran sodium sulfate. Colon transcriptome and colon lamina propria cells were evaluated in mice postcolonization by RNA-seq and ow cytometry, respectively, and T helper (T H ) 17 differentiation was assessed in vitro. Results: Mice colonized with microbiota from patients with UC low in Firmicutes had increased sensitivity to colitis compared with mice colonized with fecal or synthetic ecosystems rich in Firmicutes. Microbiota low in Firmicutes increased expression of T H 17-related genes and expansion of interleukin-17Aexpressing CD4 + cells in vivo. Supplementation with bacterial isolates belonging to the Firmicutes phylum abrogated the heightened T H 17 responses in vitro. Conclusions: A microbiota rich in Firmicutes derived from fecal samples of a healthy human donor, or assembled synthetically, downregulated colonic inammation and T H 17 pathways in mice. The results support the use of ecobiotherapy strategies, enriched in Firmicutes, for the prevention or treatment of UC. (Inamm Bowel Dis 2015;21:18831893) Key Words: ulcerative colitis, microbiota, T H 17, Firmicutes O ver the past decade, the intestinal microbiota has become one of the most studied areas in biomedical research. Gnotobiotic studies have provided key evidence on how specic bacterial strains inuence the development and maturation of host physiology and immunity, including secondary lymphoid devel- opment, antimicrobial peptide maturation, and induction of T helper (T H ) cells, such as T regulatory (T REG ) and proinam- matory T H 17 cells. 13 Components of the intestinal microbiota have differential capacity to inuence host responses. 4 For instance, experimentally dened communities, such as the altered Schaedler ora (ASF), Clostridium species, or the poly- saccharide Aexpressing Bacteroides fragilis predominantly induce T REG in the colon, whereas pathobionts Bilophila wad- sworthia and segmented lamentous bacteria are potent inducers of T H 1 and T H 17 cells, respectively. 58 Others have shown that colonization with an enterotoxigenic B. fragilis worsens exper- imental colitis in mice compared with a nontoxigenic strain. 9 Altogether, this suggests that the host-microbiota interaction sits in a continuum between mutualism and pathogenicity, and the composition of the microbiota is one factor that could potentially upset this delicate balance. Disturbed microbiota, termed dysbiosis, which is a state in which the homeostatic balance between the microbiota and the host is shifted toward a proinammatory state, has been described in a number of autoimmune and intestinal inamma- tory conditions. 10,11 It is currently accepted that ulcerative coli- tis (UC), one of the 2 forms of inammatory bowel disease (IBD), results from the development of abnormal immune re- sponses to the colonic microbiota. 12 Although therapies for UC currently aim at modulating the immune response, there has been increasing interest in microbiota-modulating therapies, Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journals Web site (www.ibdjournal.org). Received for publication February 12, 2015; Accepted March 5, 2015. From the *Farncombe Family Digestive Health Research Institute, McMaster University, Hamilton, ON, Canada; Department of Pathology, and Molecular Med- icine, McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada; and Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada. Supported by CCFC and CIHR (MOP 123282) grants to E.F.V. and E.A.-V. J.M.N. was awarded CCFC/CAG student award. The authors have no conicts of interest to disclose. Reprints: Elena F. Verdu, MD, PhD, Room HSC 3N4A, 1200 Main Street West, Hamilton, ON L8N 3Z5, Canada (e-mail: verdue@mcmaster.ca). Copyright © 2015 Crohns & Colitis Foundation of America, Inc. DOI 10.1097/MIB.0000000000000422 Published online 29 April 2015. Inamm Bowel Dis Volume 21, Number 8, August 2015 www.ibdjournal.org | 1883 Copyright © 2015 Crohns & Colitis Foundation of America, Inc. Unauthorized reproduction of this article is prohibited. 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