Enterocyte cytoskeleton changes are crucial for enhanced translocation of nonpathogenic Escherichia coli across metabolically stressed gut epithelia.
Substantial data implicate the commensal flora as triggers for the initiation of enteric inflammation or inflammatory disease relapse. We have shown that enteric epithelia under metabolic stress respond to nonpathogenic bacteria by increases in epithelial paracellular permeability and bacterial translocation. Here we assessed the structural basis of these findings. Confluent filter-grown monolayers of the human colonic T84 epithelial cell line were treated with 0.1 mM dinitrophenol (which uncouples oxidative phosphorylation) and noninvasive, nonpathogenic Escherichia coli (strain HB101, 10(6) CFU) with or without pretreatment with various pharmacological agents. At 24 h later, apoptosis, tight-junction protein expression, transepithelial resistance (TER; a marker of paracellular permeability), and bacterial internalization and translocation were assessed. Treatment with stabilizers of microtubules (i.e., colchicine), microfilaments (i.e., jasplakinolide) and clathrin-coated pit endocytosis (i.e., phenylarsine oxide) all failed to block DNP+E. coli HB101-induced reductions in TER but effectively prevented bacterial internalization and translocation. Neither the TER defect nor the enhanced bacterial translocations were a consequence of increased apoptosis. These data show that epithelial paracellular and transcellular (i.e., bacterial internalization) permeation pathways are controlled by different mechanisms. Thus, epithelia under metabolic stress increase their endocytotic activity that can result in a microtubule-, microfilament-dependent internalization and transcytosis of bacteria. We speculate that similar events in vivo would allow excess unprocessed antigen and bacteria into the mucosa and could evoke an inflammatory response by, for example, the activation of resident or recruited immune cells.
Nazli
Aisha
A
Intestinal Disease Research Programme, HSC-3N5C, McMaster University, 1200 Main Street West, Hamilton, Ontario L8N 3Z5, Canada.
Wang
Arthur
A
Steen
Oren
O
Prescott
David
D
Lu
Jun
J
Perdue
Mary H
MH
Söderholm
Johan D
JD
Sherman
Philip M
PM
McKay
Derek M
DM
D018988:Bacterial Translocation / Q000187:drug effects / Q000276:immunology*; D045744:Cell Line, Tumor; D002701:Chloramphenicol / Q000494:pharmacology; D003599:Cytoskeleton / Q000276:immunology* / Q000378:metabolism* / Q000382:microbiology; D004140:Dinitrophenols / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects / Q000276:immunology; D020895:Enterocytes / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013312:Stress, Physiological / Q000378:metabolism* / Q000382:microbiology
D004140:Dinitrophenols; D011500:Protein Synthesis Inhibitors; D002701:Chloramphenicol