Stebbings S, Munro K, Simon M A, Tannock G, Highton J, Harmsen H, Welling G, Seksik P, Dore J, Grame G, Tilsala-Timisjarvi A
Department of Rheumatology, Southmead Hospital, Westbury-on-Trym, Bristol, UK.
Rheumatology (Oxford). 2002 Dec;41(12):1395-401. doi: 10.1093/rheumatology/41.12.1395.
To determine whether differences within the complex intestinal microflora can be demonstrated between patients with ankylosing spondylitis (AS) and healthy individuals.
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.
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.
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.
确定强直性脊柱炎（AS）患者与健康个体之间是否能显示出复杂肠道微生物群的差异。
采用多种基于核酸的方法，包括变性梯度凝胶电泳（DGGE），来确定15例强直性脊柱炎患者和15例匹配对照的粪便微生物群组成。测定与肠道细菌反应的血清抗体浓度。使用生物发光测定法确定对自体肠道细菌的T细胞增殖反应。
DGGE显示每个个体粪便中有独特且稳定的细菌群落。患者和对照之间在定植谱上没有明显的特异性差异。使用基于核酸的方法对单个细菌组进行分析，结果显示肺炎克雷伯菌或普通拟杆菌在粪便定植方面没有差异。与对照组样本相比，发现AS患者粪便样本中含有硫酸盐还原菌的比例显著更高（P = 0.0004）。五分之三的患者对从其自身粪便中培养出的拟杆菌属物种表现出升高的T细胞增殖反应。患者组中与克雷伯菌或拟杆菌细胞反应的血清免疫球蛋白A（IgA）和IgM抗体浓度相对于对照组较低。
通过使用DGGE，我们证明了人类肠道微生物群的复杂性和个体性，并表明这是确定个体生物体在脊柱关节炎发病机制中可能意义的一个混杂因素。尽管如此，我们证明了AS患者粪便中硫酸盐还原菌的患病率较高。这些生物体与炎症性肠病的发病机制有关。我们还检测到AS患者对自体拟杆菌分离株可能存在免疫耐受性丧失。
