Enterocyte TLR4 mediates phagocytosis and translocation of bacteria across the intestinal barrier.
Translocation of bacteria across the intestinal barrier is important in the pathogenesis of systemic sepsis, although the mechanisms by which bacterial translocation occurs remain largely unknown. We hypothesized that bacterial translocation across the intact barrier occurs after internalization of the bacteria by enterocytes in a process resembling phagocytosis and that TLR4 is required for this process. We now show that FcgammaRIIa-transfected enterocytes can internalize IgG-opsonized erythrocytes into actin-rich cups, confirming that these enterocytes have the molecular machinery required for phagocytosis. We further show that enterocytes can internalize Escherichia coli into phagosomes, that the bacteria remain viable intracellularly, and that TLR4 is required for this process to occur. TLR4 signaling was found to be necessary and sufficient for phagocytosis by epithelial cells, because IEC-6 intestinal epithelial cells were able to internalize LPS-coated, but not uncoated, latex particles and because MD2/TLR4-transfected human endothelial kidney (HEK)-293 cells acquired the capacity to internalize E. coli, whereas nontransfected HEK-293 cells and HEK-293 cells transfected with dominant-negative TLR4 bearing a P712H mutation did not. LPS did not induce membrane ruffling or macropinocytosis in enterocytes, excluding their role in bacterial internalization. Strikingly, the internalization of Gram-negative bacteria into enterocytes in vivo and the translocation of bacteria across the intestinal epithelium to mesenteric lymph nodes were significantly greater in wild-type mice as compared with mice having mutations in TLR4. These data suggest a novel mechanism by which bacterial translocation occurs and suggest a critical role for TLR4 in the phagocytosis of bacteria by enterocytes in this process.
Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, PA 15213, USA.
Matthew D
MD
Neal
Cynthia
C
Leaphart
Ryan
R
Levy
Jose
J
Prince
Timothy R
TR
Billiar
Simon
S
Watkins
Jun
J
Li
Selma
S
Cetin
Henri
H
Ford
Alan
A
Schreiber
David J
DJ
Hackam
D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D018988:Bacterial Translocation / Q000235:genetics / Q000276:immunology*; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D020895:Enterocytes / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology* / Q000382:microbiology; D008070:Lipopolysaccharides / Q000378:metabolism; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D018345:Mice, Knockout; D010587:Phagocytosis / Q000276:immunology*; D017452:Receptors, IgG / Q000235:genetics; D012756:Sheep; D051197:Toll-Like Receptor 4 / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D015703:Antigens, CD; C080568:Fc gamma receptor IIA; D008070:Lipopolysaccharides; D017452:Receptors, IgG; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4