Riedel Christian U, Foata Francis, Goldstein Darlene R, Blum Stephanie, Eikmanns Bernhard J
Department of Microbiology and Biotechnology, University of Ulm, 89068 Ulm, Germany.
Int J Food Microbiol. 2006 Jul 1;110(1):62-8. doi: 10.1016/j.ijfoodmicro.2006.01.040. Epub 2006 May 19.
The aim of the present study was to study different strains of bifidobacteria for adhesion to Caco-2 intestinal epithelial cells (IECs) and to test for the mRNA response of these cells following interaction with bifidobacteria. Adhesion was tested at different pH conditions using model epithelia consisting of transwell cultures of fully differentiated Caco-2 cells. Microarrays were used to characterize changes in global expression profiles of Caco-2 cells co-cultured with peripheral blood mononuclear cells (PBMCs) and challenged with non-pathogenic Escherichia coli D2241 or four different strains of bifidobacteria. Furthermore, cytokine mRNA of IECs in responses to challenge with Bifidobacterium bifidum S17 or E. coli D2241 was tested in PBMC-sensitised Caco-2 cells using RT-PCR. Bifidobacteria showed strain-specific adhesion to Caco-2. Shift of apical pH from 7 to 4.5 resulted in strain-specific changes of adhesion. Global expression profiles of PBMC-sensitised Caco-2 cells revealed differential expression of a significant number of genes only after challenge with E. coli D2241 while cells were essentially unresponsive to challenge with four strains of bifidobacteria showing different adhesion properties. Using a RT-PCR approach, in the same system a similar differential expression after challenge with E. coli D2241 or B. bifidum S17 was observed for various immune markers. The presented results suggest that Caco-2 cells might be specifically unresponsive to challenge with bifidobacteria irrespective of the level of adhesion.
本研究的目的是研究不同双歧杆菌菌株对Caco-2肠上皮细胞(IECs)的黏附情况,并检测这些细胞与双歧杆菌相互作用后的mRNA反应。使用由完全分化的Caco-2细胞的Transwell培养物组成的模型上皮,在不同pH条件下测试黏附情况。利用微阵列来表征与外周血单核细胞(PBMCs)共培养并受到非致病性大肠杆菌D2241或四种不同双歧杆菌菌株攻击的Caco-2细胞的整体表达谱变化。此外,使用RT-PCR在PBMC致敏的Caco-2细胞中检测IECs对两歧双歧杆菌S17或大肠杆菌D2241攻击的细胞因子mRNA。双歧杆菌对Caco-2表现出菌株特异性黏附。顶端pH从7变为4.5导致黏附的菌株特异性变化。PBMC致敏的Caco-2细胞的整体表达谱显示,仅在受到大肠杆菌D2241攻击后大量基因存在差异表达,而细胞对四种具有不同黏附特性的双歧杆菌菌株的攻击基本无反应。使用RT-PCR方法,在同一系统中,对于各种免疫标志物,在用大肠杆菌D2241或两歧双歧杆菌S17攻击后观察到类似的差异表达。呈现的结果表明,无论黏附水平如何,Caco-2细胞可能对双歧杆菌的攻击具有特异性无反应。