Bacteroides thetaiotaomicron ameliorates colon inflammation in preclinical models of Crohn’s Disease [Bacteroides]

Analysis of the Bacteroides thetaiotaomicron(BT) transcriptome during co-culture with Caco-2 intestinal epithelial cells To identify potential bacterial protein(s) involved in the anti-inflammatory effect of BT in colitis, BT was incubated with Caco-2 human intestinal epithelial cells for 2 hours, and bacterial gene expression was assessed on a Bacteroides thetaiotaomicron VPI-5482 specific microarray. Forty-three BT genes were up-regulated by five-fold or more and of these, twenty genes encoded hypothetical proteins. Caco-2 cells cultured in 35mm dishes were treated with flagellin of Salmonella enteritidis, phorbol-12-myristate-13-acetate or interleukin 1. BT was added at 109 CFU/mL and incubated for 2h. Bacterial culture was then removed and RNA was processed for hybridization to a Bacteroides thetaiotaomicron VPI-5482 specific microarray.

多形拟杆菌（Bacteroides thetaiotaomicron, BT）与Caco-2人肠道上皮细胞共培养过程中的转录组分析：为鉴定参与BT在结肠炎中发挥抗炎作用的潜在细菌蛋白，将BT与Caco-2人肠道上皮细胞孵育2小时，随后采用多形拟杆菌VPI-5482特异性微阵列（microarray）检测细菌基因表达情况。本次分析共筛选得到43个上调幅度达5倍及以上的BT基因，其中20个基因编码假想蛋白（hypothetical protein）。将接种于35mm培养皿中的Caco-2细胞分别用肠炎沙门氏菌（Salmonella enteritidis）鞭毛蛋白、佛波醇-12-肉豆蔻酸酯-13-乙酸酯及白细胞介素1处理后，以10^9菌落形成单位/毫升（CFU/mL）的浓度加入BT并孵育2小时，随后移除细菌培养液，提取总RNA用于与多形拟杆菌VPI-5482特异性微阵列进行杂交实验。