Brown M, Hughes K R, Moossavi S, Robins A, Mahida Y R
Institute of Infection, Immunity and Inflammation, University of Nottingham, UK; Nottingham Digestive Diseases Centre, University of Nottingham, UK.
Clin Exp Immunol. 2014 Oct;178(1):28-39. doi: 10.1111/cei.12381.
The aim of our studies was to investigate the expression of Toll-like receptor (TLR)-2 and TLR-4 (and in some studies TLR-5) in myofibroblasts and small and large intestinal crypt epithelial cells from control patients and those affected by Crohn's disease and ulcerative colitis. Isolated and disaggregated crypt epithelial cells and monolayers of myofibroblasts were used for studies by reverse transcription-polymerase chain reaction (RT-PCR), real-time RT-PCR, flow cytometry, immunocytochemistry and Western blot analysis. Compared to control cells, crypt epithelial cells isolated from active ulcerative colitis and Crohn's disease colonic mucosal samples showed significantly higher expression of TLR-2 and TLR-4 transcripts and protein (on the cell surface). There was also enhanced expression of TLR-4 in crypt cells from ileal Crohn's disease. Expression of TLR-2 and TLR-4 transcripts in crypt epithelial cells isolated from inflamed mucosa of distal ulcerative colitis did not differ significantly from such cells obtained from the normal proximal colon. Crypt epithelial cells with side population characteristics (putative stem cells) also expressed transcripts and protein for TLR-2, TLR-4 and TLR-5. Colonic myofibroblast expression of these TLRs was much weaker than in crypt epithelial cells. In conclusion, enhanced TLR-2 and TLR-4 expression by crypt epithelial cells in active inflammatory bowel disease likely reflects greater ability to respond to microbial products. Results from our studies using mucosal samples from patients with distal ulcerative colitis suggest that the enhanced expression of these TLRs could be constitutive. TLR-2, TLR-4 and TLR-5 expression by stem cells imply ability to respond to distinct bacterial products.
我们研究的目的是调查Toll样受体（TLR）-2和TLR-4（在一些研究中还包括TLR-5）在来自对照患者以及克罗恩病和溃疡性结肠炎患者的肌成纤维细胞、小肠和大肠隐窝上皮细胞中的表达情况。分离和解离的隐窝上皮细胞以及肌成纤维细胞单层用于通过逆转录-聚合酶链反应（RT-PCR）、实时RT-PCR、流式细胞术、免疫细胞化学和蛋白质印迹分析进行研究。与对照细胞相比，从活动性溃疡性结肠炎和克罗恩病结肠黏膜样本中分离出的隐窝上皮细胞显示出TLR-2和TLR-4转录本及蛋白（在细胞表面）的表达显著更高。回肠克罗恩病隐窝细胞中TLR-4的表达也增强。从远端溃疡性结肠炎炎症黏膜中分离出的隐窝上皮细胞中TLR-2和TLR-4转录本的表达与从正常近端结肠获得的此类细胞相比无显著差异。具有侧群特征的隐窝上皮细胞（推测为干细胞）也表达TLR-2、TLR-4和TLR-5的转录本和蛋白。结肠肌成纤维细胞中这些TLR的表达比隐窝上皮细胞弱得多。总之，活动性炎症性肠病中隐窝上皮细胞增强的TLR-2和TLR-4表达可能反映了对微生物产物的更大反应能力。我们使用远端溃疡性结肠炎患者黏膜样本的研究结果表明，这些TLR的增强表达可能是组成性的。干细胞表达TLR-2、TLR-4和TLR-5意味着其具有对不同细菌产物作出反应的能力。
