Rheumatology 2002; 41: 1395-1401
© 2002 British Society for Rheumatology
Original Papers |
Comparison of the faecal microflora of patients with ankylosing spondylitis and controls using molecular methods of analysis
1 Department of Rheumatology, Southmead Hospital, Bristol, UK,
2 Department of Microbiology, University of Otago, Dunedin,
3 Department of Medical and Surgical Sciences, University of Otago, New Zealand,
4 Department of Medical Microbiology, University of Groningen, The Netherlands,
5 INRA, Jouy-en-Josas, France and
6 Department of Biology, University of Oulu, Finland
Objectives. 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.
KEY WORDS: Faecal microflora, Ankylosing spondylitis, Sulphate-reducing bacteria, Denaturing gradient gel electrophoresis, PCR.
Correspondence to: S. Stebbings, Musculoskeletal Directorate, Department of Rheumatology, Southmead Hospital, Westbury-on -Trym, Bristol BS10 5NB, UK.
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