A PDF file should load here. If you do not see its contents
the file may be temporarily unavailable at the journal website
or you do not have a PDF plug-in installed and enabled in your browser.
Alternatively, you can download the file locally and open with any standalone PDF reader:
http://www.ped-rheum.com/content/pdf/1546-0096-12-S1-P47.pdf
The intestinal microbiota in enthesitis-related arthitis
Alessia Paladini
0
Monica di Paola
0
Ilaria Pagnini
0
Gabriele Simonini
0
Teresa Giani
0
Paolo Lionetti
0
Rolando Cimaz
0
0
Aou Meyer - Rheumatology Department
,
Florence
,
Italy
-
From 21st European Pediatric Rheumatology (PReS) Congress
Belgrade, Serbia. 17-21 September 2014
Introduction
The role of gut inflammation and of intestinal
permeability has been extensively studied in spondyloarthropathies,
and more recently also in enthesitisrelated arthritis
(ERA). Clinical and experimental evidences have indicated
that in genetically predisposed subjects alterations of gut
microbiota may be linked to abnormal immune responses
and chronic inflammatory disorders, and oral and gut
microbiota have both been associated with RA.
Methods
We enrolled 27 consecutive children with JIA, 17 with
ERA and 10 (as controls) with oligoarticular or
polyarticular disease. The majority of cases had two separate stool
samples taken at 3 months-interval. Exclusion criteria
were presence of gastrointestinal symptoms and antibiotics
or 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 disease
duration 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 pts
with active disease.
From fecal samples, bacterial genomic DNA was
extracted and bacterial 16S rDNA, a molecular marker
used to assess microbial diversity, was amplified. All
samples were subjected to Amplified Ribosomal DNA
Restriction (ARDRA) analysis, a method that we had previously
demonstrated to be useful in assessing microbiota
dynamics on pools of genomic DNA extracted from stools
from pts with with different diseases. Four restriction
enzymes (HaeIII, AluI, MspI, RsaI) were selected to
produce restriction fragments from amplified pools of
bacterial DNA of each sample and to generate diagnostic
restriction profiles. A dendrogram, based on ARDRA
patterns, was constructed using the PHYLIP program with
1% tolerance (i.e. bands that differ by about seven
nucleotides or less are considered identical) and with the same
methodology used to compare restriction patterns from
single bacterial species isolates. The clustering of the 16S
rDNA restriction profiles on the mixed bacterial
populations allowed discriminating every sample as unique and,
in particular, to assess sample similarity and variations in
the microbiota dynamics (in the same patient) according
to a well-established methodology.
Results
The dendrogram representing the clustering of different
profiles obtained showed that samples from patients with
ERA and those oligo- or poly- JIA clustered separately.
Interestingly, HLA-B27 patients seemed to form a separate
subcluster.
Conclusion
This preliminary report shows that ERA and non-ERA
patients have different stool microbiota. We are currently
verifying the effects of drug treatment and disease activity
on these results, and at the same time performing next
generation sequencing on these samples in order to
identify specific bacterial populations.
Disclosure of interest
None declared.
(...truncated)