Prevalence of Yersinia Species in the Ileum of Crohn's Disease Patients and Controls

Guillaume Le Baut, Claire O'Brien, Paul Pavli, Maryline Roy, Philippe Seksik, Xavier Tréton, Stéphane Nancey, Nicolas Barnich, Madeleine Bezault, Claire Auzolle, Dominique Cazals-Hatem, Jérome Viala, Matthieu Allez, REMIND GROUP, Jean-Pierre Hugot, Anne Dumay, Guillaume Le Baut, Claire O'Brien, Paul Pavli, Maryline Roy, Philippe Seksik, Xavier Tréton, Stéphane Nancey, Nicolas Barnich, Madeleine Bezault, Claire Auzolle, Dominique Cazals-Hatem, Jérome Viala, Matthieu Allez, REMIND GROUP, Jean-Pierre Hugot, Anne Dumay

Abstract

Yersinia are common contaminants of food products, but their prevalence in the human gut is poorly documented. Yersinia have been implicated in Crohn's Disease (CD, an inflammatory bowel disease) however their role in CD is controversial. We performed highly sensitive PCR assays of specific sequences for the gyrB gene of Y. aldovae, Y. bercovieri, Y. enterocolitica, Y. intermedia, Y. mollaretii and the inv gene of Y. pseudotuberculosis. We analyzed a total of 470 ileal samples taken from 338 participants (262 CD patients and 76 controls) belonging to three independent cohorts. All patients and controls were phenotyped and genotyped for the main CD susceptibility variants: NOD2, ATG16L1, and IRGM. Yersinia were found in 7.7% of ileal samples (respectively 7.9 and 7.6% in controls and CD patients) corresponding to 10% of participants (respectively 11.8 and 9.5% in controls and CD patients). Y. enterocolitica, Y. pseudotuberculosis and Y. intermedia were the most frequently identified species. The bacteria were more frequent in resected specimens, lymph nodes and Peyer's patches. Yersinia were no more likely to be detected in CD tissues than tissues from inflammatory and non-inflammatory controls. CD patients treated with immunosuppressants were less likely to be Yersinia carriers. In conclusion, this work shows that Yersinia species are frequently found at low levels in the human ileum in health and disease. The role of Yersinia species in this ecosystem should now be explored.

Keywords: Crohn's disease; gut microbiota; ileal mucosa; innate immunity; molecular test; mucosal immune system; yersinia.

Figures

Figure 1
Figure 1
Validation of the PCR methods. (A) Comparison of GyrB sequences between different Yersinia species and E.coli. In red are indicated the nucleic acids present in all species. In blue is indicated the consensus sequence (Corpet, 1998). YA, Yersinia aldovae; YB, Yersinia bercovieri; YE, Yersinia enterocolitica; YI, Yersinia Intermedia; YM, Yersinia Mollaretii. (B) Specificity of the PCR methods developed for Y. frederiksenii (GyrB). DNAs from several species of Yersinia were amplified with the PCR technique and loaded on an agarose gel. A band indicates the presence of the specific PCR product specific to Y. frederiksenii. (C) Sensitivity of the PCR method developed for Y. enterocolitica (GyrB). The quantity of amplified DNA is expressed as genome equivalents. The method was highly sensitive when two cycles of 35 amplifications were performed.

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