title: Exposure of intestinal epithelial cell HT29 to bile acids and ammonia enhances Mac-1-mediated neutrophil adhesion.
abstract: OBJECTIVE
  Bile acids and ammonia (NH3/NH4+) are cytotoxic metabolic products, which are known to influence intestinal epithelial functions such as colonic chloride secretion. In this study, we have investigated the effect of bile acids and ammonia on neutrophil-intestinal epithelial adhesive interaction.
  
  
  METHODS
  Confluently cultured HT29 cells were treated with bile acids or ammonium chloride. Then, 51Cr-labeled neutrophils were added to HT29 cell monolayers, and neutrophil adhesion was assessed by gamma-scintillation counting.
  
  
  RESULTS
  Treatment of HT29 cells with 0.1 mM CDCA (chenodeoxycholic acid), DCA (deoxycholic acid), CA (cholic acid) or ammonium chloride but not UDCA (ursodeoxycholic acid) for 30 min increased neutrophil adhesion about 4-folds (p<0.01). The increased adhesion was inhibited 82-91% by 10 microg/ml anti-CD11b and anti-CD18 mAbs (p<0.01), but not by anti-CD11a and anti-CD54 (ICAM-1) mAbs. Interestingly, flow cytometric analysis revealed that ICAM-1 expression on HT29 cells was not changed by bile acid- or ammonia-treatment. In addition, the increased adhesion was inhibited about 65 % by proteinase K-treatment (10 microg/ml, 1 min, p<0.05) but not cycloheximide-treatment (1 microg/ml, 30 min) of HT29 cells.
  
  
  CONCLUSIONS
  These observations indicate that exposure of HT29 cells to bile acids or ammonia induces CD11b/CD18 (Mac-1) dependent- but CD11a/CD18 (LFA-1) independent-neutrophil adhesion to intestinal epithelial cells, and ICAM-1 is unlikely involved in the interactions. Furthermore, epithelial ligand(s) for neutrophils are protein molecule(s) which are expressed on the cell surface independent of protein synthesis.
authors.affiliation: Department of Biochemistry, Juntendo University, School of Medicine, Tokyo, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Miyata
authors.forename: K
authors.initials: K
authors.lastname: Iwabuchi
authors.forename: S
authors.initials: S
authors.lastname: Watanabe
authors.forename: N
authors.initials: N
authors.lastname: Sato
authors.forename: I
authors.initials: I
authors.lastname: Nagaoka
mesh_terms: D000641:Ammonia / Q000494:pharmacology*; D000963:Antimetabolites / Q000494:pharmacology; D001647:Bile Acids and Salts / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D005434:Flow Cytometry; D006026:Glycoside Hydrolases / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D015151:Immunoblotting; D007202:Indicators and Reagents; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000166:cytology / Q000187:drug effects; D016177:Macrophage-1 Antigen / Q000502:physiology*; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D000963:Antimetabolites; D001647:Bile Acids and Salts; D007202:Indicators and Reagents; D016177:Macrophage-1 Antigen; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D000641:Ammonia; D006026:Glycoside Hydrolases