Colonic epithelial hPepT1 expression occurs in inflammatory bowel disease: transport of bacterial peptides influences expression of MHC class 1 molecules

Gastroenterology. 2001 Jun;120(7):1666-79. doi: 10.1053/gast.2001.24845.

Abstract

Background & aims: hPepT1 is an intestinal epithelial apical membrane transporter responsible for uptake of di/tripeptides (including bacterial derived proinflammatory n-formyl peptides). hPepT1 expression normally has a strict axial gradient-highest in the proximal small intestine with no expression in the colon.

Methods: Small intestinal-like cells (Caco2-BBE), and colonic-like cells (HT29-Cl.19A), and colonic mucosa from diseased and control patients were used in the present study.

Results: hPepT1 expression occurs aberrantly in the colon with chronic ulcerative colitis (6 patients) and Crohn's disease (4 patients), but not in normal colon (4 patients) or colon with microscopic colitis (4 patients). To model expression of hPepT1 by colonic-like cells in inflamed states, we stably transfected HT29-Cl.19A cells with a modified hPepT1 tagged on the N-terminus with green fluorescence protein. Analysis of transfected cells revealed that: GFP-hPepT1 protein, like the natural protein, is targeted to the apical plasma membrane. In addition, the tagged protein retains the capability of di/tripeptide absorption, and the expression of the tagged protein by HT29-Cl.19A cells permits absorption of N-formyl-methionyl-leucyl-phenylalanine (fMLP), as occurs in hPepT1 expressing Caco2-BBE cells. fMLP uptake by colonic cells expressing GFP-hPepT1 specifically enhances major histocompatibility complex class I surface expression.

Conclusions: These data collectively indicate that, in some states of chronic inflammation, hPepT1 may be anomolously expressed in the colon. Further, transport of fMLP by hPepT1 potentially stimulates expression of key accessory immune molecule, MHC-1.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / metabolism*
  • Biological Transport
  • Caco-2 Cells
  • Carrier Proteins / analysis
  • Carrier Proteins / physiology*
  • Colitis / metabolism
  • Colon / chemistry
  • HT29 Cells
  • Histocompatibility Antigens Class I / analysis*
  • Humans
  • Inflammatory Bowel Diseases / metabolism*
  • Intestine, Small / chemistry
  • Molecular Sequence Data
  • N-Formylmethionine Leucyl-Phenylalanine / metabolism*
  • Peptide Transporter 1
  • Symporters*

Substances

  • Bacterial Proteins
  • Carrier Proteins
  • Histocompatibility Antigens Class I
  • Peptide Transporter 1
  • SLC15A1 protein, human
  • Symporters
  • N-Formylmethionine Leucyl-Phenylalanine