Core 2 Mucin-Type O-Glycan Is Related to EPEC and EHEC O157:H7 Adherence to Human Colon Carcinoma HT-29 Epithelial Cells
Core 2 Mucin-Type O-Glycan Is Related to EPEC and EHEC O157:H7 Adherence to Human Colon Carcinoma HT-29 Epithelial Cells
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DOI：
10.1007/s10620-015-3548-5
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发表时间：
2015-02
影响因子：
3.1
通讯作者：
Jun Ye;Lili Song;Yun Liu;Q. Pan;Xiaoli Zhong;Shan-shan Li;Yangyang Shang;Yin Tian;Yonghong He;Lei Chen;Wensheng Chen;Zhi-hong Peng;Rong-quan Wang
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作者：
Jun Ye;Lili Song;Yun Liu;Q. Pan;Xiaoli Zhong;Shan-shan Li;Yangyang Shang;Yin Tian;Yonghong He;Lei Chen;Wensheng Chen;Zhi-hong Peng;Rong-quan Wang
背景和目的宿主糖基化在EPEC和EHEC O 157：H7相互作用中的作用尚不清楚;本研究检测了O-聚糖是否参与EPEC和EHEC O 157：H7对HT-29细胞的粘附。方法采用粘附细菌和宿主细胞的直接共染色、粘附细菌平板和/或粘附GFP标记细菌的直接荧光观察来确定细菌对培养细胞的粘附。H7对HT-29-Gal和HT-29细胞的粘附表明，HT-29细胞的分化导致EPEC和EHEC O 157：H7的粘附减少。用苄基-α-GalNAc处理EPEC和EHEC O 157：H7，可抑制O-glycan的合成，从而降低EPEC和EHEC O 157：H7的粘附能力。与HT-29和shRNA-Ctr/HT-29细胞相比，C2 GnT 2敲低诱导的核心2 O-聚糖缺陷HT-29细胞在C2 GnT 2-sh 2/HT-29细胞中的EPEC和EHEC O 157：H7粘附显著降低。在苄基-α-GalNAc处理的HT-29细胞中MUC 2表达显著降低，但在C2 GnT 2缺陷的HT-29细胞中无变化。EPEC或EHEC O 157：H7感染C2 GnT 2缺陷的HT-29细胞后，上皮屏障功能恶化。EPEC或EHEC O 157：H7感染shRNA-Ctr/HT-29和C2 GnT 2-sh 2/HT-29细胞后，occludin的表达在细胞表面呈不连续的密集分布，而对照细胞呈连续分布。这些数据表明，EPEC和肠出血性大肠杆菌O 157：H7粘附HT-29细胞是相关的粘蛋白型核心2 O-glycan.ConclusionsThis研究提供的概念，对设计的碳水化合物依赖性抑制EPEC和肠出血性大肠杆菌O 157：H7粘附到人肠上皮细胞。
Background and AimThe roles of host glycosylation in interactions with EPEC and EHEC O157:H7 are largely unclear; this study examined whetherO-glycans are involved in EPEC and EHEC O157:H7 adherence to HT-29 cells.MethodsBacterial adherence to the cultured cells was determined using the direct co-staining of adherent bacteria and host cells, the adherent bacteria plating, and/or the direct fluorescent observation of the adherent GFP-labeled bacteria.ResultsA comparison of the adherence of EPEC and EHEC O157:H7 to HT-29-Gal and HT-29 cells indicated that the differentiation of HT-29 cells led to a reduction in the adherence of EPEC and EHEC O157:H7. EPEC and EHEC O157:H7 adhesion decreased after the abrogation ofO-glycan biosynthesis mediated by benzyl-α-GalNAc treatment. Core 2O-glycan-deficient HT-29 cells induced by C2GnT2 knockdown had a significant reduction in EPEC and EHEC O157:H7 adhesion in C2GnT2-sh2/HT-29 cells compared with HT-29 and shRNA-Ctr/HT-29 cells. MUC2 expression in benzyl-α-GalNAc-treated HT-29 cells was significantly reduced but unchanged in C2GnT2-deficient HT-29 cells. EPEC or EHEC O157:H7 infection in C2GnT2-deficient HT-29 cells deteriorated the epithelial barrier function. The occludin expression in the shRNA-Ctr/HT-29 and C2GnT2-sh2/HT-29 cells after infection with EPEC or EHEC O157:H7 was pyknic and discontinuous at the cell surface compared with its continuous distribution of control cells. These data indicate that EPEC and EHEC O157:H7 adherence to HT-29 cells is related to mucin-type core 2O-glycan.ConclusionsThis study provides the concepts toward the design of carbohydrate-dependent inhibition of EPEC and EHEC O157:H7 adhesion to human intestinal epithelial cells.