Toll-like receptor 2 activation by β2→1-fructans protects barrier function of T84 human intestinal epithelial cells in a chain length-dependent manner.
Dietary fiber intake is associated with lower incidence and mortality from disease, but the underlying mechanisms of these protective effects are unclear. We hypothesized that β2→1-fructan dietary fibers confer protection on intestinal epithelial cell barrier function via Toll-like receptor 2 (TLR2), and we studied whether β2→1-fructan chain-length differences affect this process. T84 human intestinal epithelial cell monolayers were incubated with 4 β2→1-fructan formulations of different chain-length compositions and were stimulated with the proinflammatory phorbol 12-myristate 13-acetate (PMA). Transepithelial electrical resistance (TEER) was analyzed by electric cell substrate impedance sensing (ECIS) as a measure for tight junction-mediated barrier function. To confirm TLR2 involvement in barrier modulation by β2→1-fructans, ECIS experiments were repeated using TLR2 blocking antibody. After preincubation of T84 cells with short-chain β2→1-fructans, the decrease in TEER as induced by PMA (62.3 ± 5.2%, P < 0.001) was strongly attenuated (15.2 ± 8.8%, P < 0.01). However, when PMA was applied first, no effect on recovery was observed during addition of the fructans. By blocking TLR2 on the T84 cells, the protective effect of short-chain β2→1-fructans was substantially inhibited. Stimulation of human embryonic kidney human TLR2 reporter cells with β2→1-fructans induced activation of nuclear factor kappa-light-chain-enhancer of activated B cells, confirming that β2→1-fructans are specific ligands for TLR2. To conclude, β2→1-fructans exert time-dependent and chain length-dependent protective effects on the T84 intestinal epithelial cell barrier mediated via TLR2. These results suggest that TLR2 located on intestinal epithelial cells could be a target of β2→1-fructan-mediated health effects.
Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands; L.M.Vogt@umcg.nl.
Leonie M
LM
Vogt
Sensus, Roosendaal, The Netherlands;
Diederick
D
Meyer
Cosun Food Technology Centre, Roosendaal, The Netherlands;
Gerdie
G
Pullens
Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands;
Marijke M
MM
Faas
TNO Quality of Life, Department of Biosciences, Zeist, The Netherlands; and.
Koen
K
Venema
Laboratory of Food Chemistry, Wageningen University, Wageningen, The Netherlands.
Uttara
U
Ramasamy
Laboratory of Food Chemistry, Wageningen University, Wageningen, The Netherlands.
Henk A
HA
Schols
Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands;
Paul
P
de Vos
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D019138:Antibodies, Blocking / Q000494:pharmacology; D002460:Cell Line; D003106:Colon / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D004075:Diglycerides / Q000494:pharmacology; D005630:Fructans / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D005765:Gastrointestinal Agents / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D007668:Kidney / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008024:Ligands; D049990:Membrane Transport Modulators / Q000037:antagonists & inhibitors / Q000633:toxicity; D015394:Molecular Structure; D016328:NF-kappa B / Q000819:agonists / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D056692:Prebiotics / Q000032:analysis; D020011:Protective Agents / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000633:toxicity; D019108:Tight Junctions / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000819:agonists / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D019138:Antibodies, Blocking; D004075:Diglycerides; C460864:FSL-1 lipoprotein, synthetic; D005630:Fructans; D005765:Gastrointestinal Agents; D008024:Ligands; D049990:Membrane Transport Modulators; D016328:NF-kappa B; D009842:Oligopeptides; D056692:Prebiotics; D020011:Protective Agents; D011994:Recombinant Proteins; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; D018808:Transcription Factor AP-1; C016658:phorbolol myristate acetate; D013755:Tetradecanoylphorbol Acetate