Cimaz, R.

Di Paola, M

Giani, T

Lionetti, P

Pagnini, I

Paladini, Adriano

Simonini, G

English

Florence Research

POSTER PRESENTATION Open AccessThe intestinal microbiota in enthesitis-relatedarthitisAlessia Paladini, Monica di Paola, Ilaria Pagnini, Gabriele Simonini, Teresa Giani, Paolo Lionetti, Rolando Cimaz*From 21st European Pediatric Rheumatology (PReS) CongressBelgrade, Serbia. 17-21 September 2014IntroductionThe role of gut inflammation and of intestinal permeabil-ity has been extensively studied in spondyloarthropathies,and more recently also in enthesitis–related arthritis(ERA). Clinical and experimental evidences have indicatedthat in genetically predisposed subjects alterations of gutmicrobiota may be linked to abnormal immune responsesand chronic inflammatory disorders, and oral and gutmicrobiota have both been associated with RA.ObjectivesIn the present study, we have assessed microbial diver-sity in the gut of patients with ERA compared withother JIA diagnoses.MethodsWe enrolled 27 consecutive children with JIA, 17 withERA and 10 (as controls) with oligoarticular or polyarticu-lar disease. The majority of cases had two separate stoolsamples taken at 3 months-interval. Exclusion criteriawere presence of gastrointestinal symptoms and antibioticsor probiotics consumption in the month prior to sampling.Median age at sampling was 13 and 9 years, respectively,in the two groups (ranges, 8-17, and 6-14). Median diseaseduration was 9 y in the first group (range, 6-14) and 2 y(1-6) in the second group. In each group there were 4 ptswith active disease.From fecal samples, bacterial genomic DNA wasextracted and bacterial 16S rDNA, a molecular markerused to assess microbial diversity, was amplified. All sam-ples were subjected to Amplified Ribosomal DNA Restric-tion (ARDRA) analysis, a method that we had previouslydemonstrated to be useful in assessing microbiotadynamics on pools of genomic DNA extracted from stoolsfrom pts with with different diseases. Four restrictionenzymes (HaeIII, AluI, MspI, RsaI) were selected to pro-duce restriction fragments from amplified pools of bacter-ial DNA of each sample and to generate diagnosticrestriction profiles. A dendrogram, based on ARDRA pat-terns, was constructed using the PHYLIP program with1% tolerance (i.e. bands that differ by about seven nucleo-tides or less are considered identical) and with the samemethodology used to compare restriction patterns fromsingle bacterial species isolates. The clustering of the 16SrDNA restriction profiles on the mixed bacterial popula-tions allowed discriminating every sample as unique and,in particular, to assess sample similarity and variations inthe microbiota dynamics (in the same patient) accordingto a well-established methodology.ResultsThe dendrogram representing the clustering of differentprofiles obtained showed that samples from patients withERA and those oligo- or poly- JIA clustered separately.Interestingly, HLA-B27 patients seemed to form a separatesubcluster.ConclusionThis preliminary report shows that ERA and non-ERApatients have different stool microbiota. We are currentlyverifying the effects of drug treatment and disease activityon these results, and at the same time performing nextgeneration sequencing on these samples in order to iden-tify specific bacterial populations.Disclosure of interestNone declared.Published: 17 September 2014Aou Meyer - Rheumatology Department, Florence, ItalyPaladini et al. Pediatric Rheumatology 2014, 12(Suppl 1):P47http://www.ped-rheum.com/content/12/S1/P47© 2014 Paladini et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.doi:10.1186/1546-0096-12-S1-P47Cite this article as: Paladini et al.: The intestinal microbiota in enthesitis-related arthitis. Pediatric Rheumatology 2014 12(Suppl 1):P47.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color figure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionSubmit your manuscript at www.biomedcentral.com/submitPaladini et al. Pediatric Rheumatology 2014, 12(Suppl 1):P47http://www.ped-rheum.com/content/12/S1/P47Page 2 of 2

The intestinal microbiota in enthesitis-related arthitis

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The intestinal microbiota in enthesitis-related arthitis

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