Oxazolone-Induced Enterocolitis in Zebrafish Depends on the Composition of the Intestinal Microbiota

Background & AimsThe pathogenesis of inflammatory bowel disease involves dysfunctional mucosal immune responses to commensal bacteria in genetically predisposed hosts. Interactions between host cells and bacteria are complicated, making it a challenge to assess their relative contribution to intestinal pathology. We developed a zebrafish model of enterocolitis to study these interactions.MethodsEnterocolitis was induced by intrarectal administration of the hapten oxazolone in adult wild-type and myeloperoxidase-reporter transgenic zebrafish in the presence or absence of antibiotics. Intestinal inflammation was evaluated by histological and flow cytometry analyses and cytokine profiling with quantitative real-time polymerase chain reaction. Changes in the composition of the intestinal microbiota following antibiotic administration were assessed by 16SrRNA sequencing and bacterial load was quantified by culture on nonselective media (colony-forming units).ResultsIn zebrafish, the infiltrate and severity of oxazolone-induced enterocolitis are influenced by the composition of the microbiota. Inflammation is characterized by granulocyte influx; epithelial damage; goblet cell depletion; and increased expression of interleukin-1β, tumor necrosis factor–α, and interleukin-10. Zebrafish given vancomycin had bacterial populations dominated by Fusobacteria and reduced enterocolitis scores, intestinal damage, and percentages of infiltrating neutrophils and eosinophils. In contrast, zebrafish given colistin sulphate had a predominance of proteobacteria and reduced eosinophil and lymphocyte infiltration, but enterocolitis scores were not reduced.ConclusionsIn zebrafish with oxazolone-induced enterocolitis, components of the intestinal microbiota affect the severity and composition of the intestinal infiltrate. The pathogenesis of inflammatory bowel disease involves dysfunctional mucosal immune responses to commensal bacteria in genetically predisposed hosts. Interactions between host cells and bacteria are complicated, making it a challenge to assess their relative contribution to intestinal pathology. We developed a zebrafish model of enterocolitis to study these interactions. Enterocolitis was induced by intrarectal administration of the hapten oxazolone in adult wild-type and myeloperoxidase-reporter transgenic zebrafish in the presence or absence of antibiotics. Intestinal inflammation was evaluated by histological and flow cytometry analyses and cytokine profiling with quantitative real-time polymerase chain reaction. Changes in the composition of the intestinal microbiota following antibiotic administration were assessed by 16SrRNA sequencing and bacterial load was quantified by culture on nonselective media (colony-forming units). In zebrafish, the infiltrate and severity of oxazolone-induced enterocolitis are influenced by the composition of the microbiota. Inflammation is characterized by granulocyte influx; epithelial damage; goblet cell depletion; and increased expression of interleukin-1β, tumor necrosis factor–α, and interleukin-10. Zebrafish given vancomycin had bacterial populations dominated by Fusobacteria and reduced enterocolitis scores, intestinal damage, and percentages of infiltrating neutrophils and eosinophils. In contrast, zebrafish given colistin sulphate had a predominance of proteobacteria and reduced eosinophil and lymphocyte infiltration, but enterocolitis scores were not reduced. In zebrafish with oxazolone-induced enterocolitis, components of the intestinal microbiota affect the severity and composition of the intestinal infiltrate.