Bibiloni Rodrigo, Mangold Marco, Madsen Karen L, Fedorak Richard N, Tannock Gerald W
Department of Medicine, University of Alberta, Edmonton, Canada.
J Med Microbiol. 2006 Aug;55(Pt 8):1141-1149. doi: 10.1099/jmm.0.46498-0.
The bacterial community (microbiota) that inhabits the gut of humans appears to be an important source of antigens that drive the chronic immunological processes characteristic of Crohn's disease (CD) and ulcerative colitis (UC). Most of the members of the microbiota have not yet been cultured, but nucleic-acid-based methods of detection and enumeration can provide information about the community. This investigation used these methods to obtain information about the bacteria associated with mucosal surfaces in the gut of 20 CD and 15 UC patients. Biopsies were collected from inflamed and non-inflamed sites in the intestines of newly diagnosed, untreated patients. Biopsies were also collected from several intestinal sites of 14 healthy subjects. The bacterial collections associated with the biopsies were analysed by generating PCR/denaturing gradient gel electrophoresis (DGGE) profiles, the preparation of 16S rRNA gene clone libraries, and qualitative PCR to detect specific groups of bacteria. The total numbers of bacteria associated with the biopsies were determined by real-time quantitative PCR. DGGE profiles generated from the terminal ileum and various colonic regions were characteristic of each individual but differed between subjects. DGGE profiles and 16S rRNA gene libraries showed that the bacteria associated with inflamed and non-inflamed tissues did not differ. UC patients had more bacteria associated with biopsies than did CD patients (P<0.01). Statistical analysis of the composition of 16S rRNA gene libraries showed that the bacterial collections in UC and CD patients differed (P<0.05). Unclassified members of the phylum Bacteroidetes were more prevalent in CD than in UC patients. Therefore, the types and numbers of bacteria associated with biopsy samples were distinctly different for UC and CD patients. The observations made in this study should permit targeting of specific bacteriological abnormalities in investigations of the pathogenesis of inflammatory bowel diseases and provide targets for medical interventions.
栖息于人类肠道的细菌群落(微生物群)似乎是驱动克罗恩病(CD)和溃疡性结肠炎(UC)所特有的慢性免疫过程的重要抗原来源。微生物群的大多数成员尚未得到培养,但基于核酸的检测和计数方法能够提供有关该群落的信息。本研究采用这些方法获取了20例CD患者和15例UC患者肠道黏膜表面相关细菌的信息。从新诊断的、未经治疗的患者肠道的炎症部位和非炎症部位采集活检样本。还从14名健康受试者的多个肠道部位采集了活检样本。通过生成PCR/变性梯度凝胶电泳(DGGE)图谱、制备16S rRNA基因克隆文库以及用于检测特定细菌群的定性PCR,对与活检样本相关的细菌菌群进行分析。通过实时定量PCR确定与活检样本相关的细菌总数。从回肠末端和各个结肠区域生成的DGGE图谱具有个体特征,但在不同受试者之间存在差异。DGGE图谱和16S rRNA基因文库显示,与炎症组织和非炎症组织相关的细菌并无差异。UC患者活检样本中的细菌比CD患者更多(P<0.01)。对16S rRNA基因文库组成的统计分析表明,UC患者和CD患者的细菌菌群存在差异(P<0.05)。拟杆菌门的未分类成员在CD患者中比在UC患者中更为普遍。因此,UC患者和CD患者活检样本相关细菌的类型和数量明显不同。本研究中的观察结果应有助于在炎症性肠病发病机制的研究中针对特定的细菌学异常情况,并为医学干预提供靶点。