Comparison of the faecal microflora of patients with ankylosing spondylitis and controls using molecular methods of analysis.
OBJECTIVE
To determine whether differences within the complex intestinal microflora can be demonstrated between patients with ankylosing spondylitis (AS) and healthy individuals.


METHODS
The composition of the faecal microflora of 15 ankylosing spondylitis patients and 15 matched controls was determined using a variety of nucleic acid-based methods, including denaturing gradient gel electrophoresis (DGGE). Concentrations of serum antibodies reactive with intestinal bacteria were determined. T-cell proliferation responses to autologous intestinal bacteria were determined using a bioluminescent assay.


RESULTS
DGGE demonstrated a unique and stable bacterial community in the faeces of each individual. No specific differences in colonization profiles were discernible between patients and controls. Analysis of individual bacterial groups using nucleic acid-based methods showed no differences in faecal colonization with Klebsiella pneumoniae or Bacteroides vulgatus. A significantly higher proportion of faecal samples from AS patients were found to contain sulphate-reducing bacteria compared with samples from controls (P=0.0004). Three out of five patients showed elevated T-cell proliferation responses to Bacteroides species cultured from their own faeces. The concentrations of serum immunoglobulin A (IgA) and IgM antibodies reactive with klebsiella or bacteroides cells were lower in the patient group relative to the controls.


CONCLUSIONS
By using DGGE, we have demonstrated the complexity and individuality of the human intestinal microflora and shown that this is a confounding factor in determining the possible significance of individual organisms in the pathogenesis of spondyloarthritis. Nevertheless, we demonstrated a higher prevalence of sulphate-reducing bacteria in the faeces of patients with AS. These organisms have been implicated in the pathogenesis of inflammatory bowel disease. We also detected a possible loss of immunological tolerance to autologous Bacteroides isolates in patients with AS.
Stebbings
S
S
Department of Rheumatology, Southmead Hospital, Westbury-on-Trym, Bristol, UK.
Munro
K
K
Simon
M A
MA
Tannock
G
G
Highton
J
J
Harmsen
H
H
Welling
G
G
Seksik
P
P
Dore
J
J
Grame
G
G
Tilsala-Timisjarvi
A
A
D001439:Bacteroides / Q000235:genetics / Q000276:immunology; D016022:Case-Control Studies; D004269:DNA, Bacterial / Q000032:analysis*; D005243:Feces / Q000382:microbiology*; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007709:Klebsiella / Q000235:genetics; D008213:Lymphocyte Activation; D016133:Polymerase Chain Reaction / Q000379:methods; D013167:Spondylitis, Ankylosing / Q000276:immunology / Q000382:microbiology*; D016999:Sulfur-Reducing Bacteria / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology
D004269:DNA, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M