title: Core 2 Mucin-Type O-Glycan Is Related to EPEC and EHEC O157:H7 Adherence to Human Colon Carcinoma HT-29 Epithelial Cells.
abstract: OBJECTIVE
  The roles of host glycosylation in interactions with EPEC and EHEC O157:H7 are largely unclear; this study examined whether O-glycans are involved in EPEC and EHEC O157:H7 adherence to HT-29 cells.
  
  
  METHODS
  Bacterial 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.
  
  
  RESULTS
  A 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 of O-glycan biosynthesis mediated by benzyl-α-GalNAc treatment. Core 2 O-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 2 O-glycan.
  
  
  CONCLUSIONS
  This study provides the concepts toward the design of carbohydrate-dependent inhibition of EPEC and EHEC O157:H7 adhesion to human intestinal epithelial cells.
authors.affiliation: Department of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Ye
authors.forename: Lili
authors.initials: L
authors.lastname: Song
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.forename: Qiong
authors.initials: Q
authors.lastname: Pan
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhong
authors.forename: Shanshan
authors.initials: S
authors.lastname: Li
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Shang
authors.forename: Yin
authors.initials: Y
authors.lastname: Tian
authors.forename: Yonghong
authors.initials: Y
authors.lastname: He
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Wensheng
authors.initials: W
authors.lastname: Chen
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Peng
authors.forename: Rongquan
authors.initials: R
authors.lastname: Wang
mesh_terms: D000116:Acetylgalactosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000906:Antibodies; D001422:Bacterial Adhesion / Q000502:physiology*; D001593:Benzyl Compounds / Q000494:pharmacology; D054324:Enterohemorrhagic Escherichia coli / Q000235:genetics / Q000378:metabolism*; D054308:Enteropathogenic Escherichia coli / Q000235:genetics / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D055262:Mucin-2 / Q000235:genetics / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference
chemical_list: D000906:Antibodies; D001593:Benzyl Compounds; C524625:MUC2 protein, human; D055262:Mucin-2; C080739:benzyl-alpha-N-acetylgalactosamine; D017351:N-Acetylglucosaminyltransferases; C117361:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-N-acetylglucosaminyltransferase 3; D000116:Acetylgalactosamine