Lactobacillus rhamnosus GG attenuates interferon-γ and tumour necrosis factor-α-induced barrier dysfunction and pro-inflammatory signalling

Microbiology (United Kingdom) - Tập 156 Số 11 - Trang 3288-3297 - 2010
Kevin A. Donato1,2, Mélanie G. Gareau1, Yu Jing Jenny Wang1, Philip M. Sherman1,2
1Cell Biology Program, Research Institute, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada
2Department of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada

Tóm tắt

The intestinal epithelium forms a protective barrier against luminal contents and the external environment, mediated via intercellular tight junctions (TJs). The TJ can be disrupted via cell signalling induced by either enteric pathogens or pro-inflammatory cytokines, thereby contributing to various intestinal disorders ranging from acute infectious diarrhoea to chronic inflammatory bowel diseases. Probiotics, such asLactobacillus rhamnosusGG (LGG), are reported to confer beneficial effects on epithelial cells, including antagonizing infections and reducing overt pro-inflammatory responses, but the underlying mechanisms of these observed effects require further characterization. We hypothesized that probiotics preserve barrier function by interfering with pro-inflammatory cytokine signalling. Caco-2bbe cells were seeded into Transwells to attain polarized monolayers with intercellular TJs. Monolayers were inoculated apically with the probiotic LGG 3 h prior to the addition of IFN-γ(100 ng ml−1) to the basolateral medium overnight. The monolayers were then placed in fresh basal medium±TNF-α(10 ng ml−1) and transepithelial electrical resistance (TER) measurements were taken over the time-course of TNF-αstimulation. To complement the TER findings, cells were processed for zona occludens-1 (ZO-1) immunofluorescence staining. As a measure of TNF-αdownstream signalling, cells were immunofluorescently stained for NF-κB p65 subunit and CXCL-8 mRNA was quantified by qRT-PCR. Basal cell culture medium was collected after overnight TNF-αstimulation to measure secreted chemokines, including CXCL-8 (interleukin-8) and CCL-11 (eotaxin). Following LGG inoculation, IFN-γpriming and 24 h TNF-αstimulation, epithelial cells maintained TER and ZO-1 distribution. LGG diminished the nuclear translocation of p65, demonstrated by both immunofluorescence and CXCL-8 mRNA expression. CXCL-8 and CCL-11 protein levels were decreased in LGG-inoculated, cytokine-challenged cells. These findings indicate that LGG alleviates the effects of pro-inflammatory cytokines on epithelial barrier integrity and inflammation, mediated, at least in part, through inhibition of NF-κB signalling.

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Tài liệu tham khảo

Ahrens, 2008, Intestinal macrophage/epithelial cell-derived CCL11/eotaxin-1 mediates eosinophil recruitment and function in pediatric ulcerative colitis, J Immunol, 181, 7390, 10.4049/jimmunol.181.10.7390

Ait-Belgnaoui, 2006, Lactobacillus farciminis treatment suppresses stress induced visceral hypersensitivity: a possible action through interaction with epithelial cell cytoskeleton contraction, Gut, 55, 1090, 10.1136/gut.2005.084194

Baldassarre, 2010, Lactobacillus GG improves recovery in infants with blood in the stools and presumptive allergic colitis compared with extensively hydrolyzed formula alone, J Pediatr, 156, 397, 10.1016/j.jpeds.2009.09.012

Basuroy, 2006, MAPK interacts with occludin and mediates EGF-induced prevention of tight junction disruption by hydrogen peroxide, Biochem J, 393, 69, 10.1042/BJ20050959

Blanchard, 2009, Chemotactic factors associated with eosinophilic gastrointestinal diseases, Immunol Allergy Clin North Am, 29, 141, 10.1016/j.iac.2008.10.002

Bruewer, 2003, Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms, J Immunol, 171, 6164, 10.4049/jimmunol.171.11.6164

Chen, 2001, Increased serum levels of eotaxin in patients with inflammatory bowel disease, Scand J Gastroenterol, 36, 515, 10.1080/003655201750153377

Choi, 2008, Effect of Lactobacillus GG and conditioned media on IL-1 β-induced IL-8 production in Caco-2 cells, Scand J Gastroenterol, 43, 938, 10.1080/00365520801965373

Donato, 2008, Escherichia albertii and Hafnia alvei are candidate enteric pathogens with divergent effects on intercellular tight junctions, Microb Pathog, 45, 377, 10.1016/j.micpath.2008.09.004

Ewaschuk, 2008, Secreted bioactive factors from Bifidobacterium infantis enhance epithelial cell barrier function, Am J Physiol Gastrointest Liver Physiol, 295, G1025, 10.1152/ajpgi.90227.2008

Fish, 1999, Synergistic effects of interferon γ and tumour necrosis factor α on T84 cell function, Gut, 45, 191, 10.1136/gut.45.2.191

Johnson-Henry, 2005, Amelioration of the effects of Citrobacter rodentium infection in mice by pretreatment with probiotics, J Infect Dis, 191, 2106, 10.1086/430318

Johnson-Henry, 2007, Surface-layer protein extracts from Lactobacillus helveticus inhibit enterohaemorrhagic Escherichia coli O157 : H7 adhesion to epithelial cells, Cell Microbiol, 9, 356, 10.1111/j.1462-5822.2006.00791.x

Johnson-Henry, 2008, Lactobacillus rhamnosus strain GG prevents enterohemorrhagic Escherichia coli O157 : H7-induced changes in epithelial barrier function, Infect Immun, 76, 1340, 10.1128/IAI.00778-07

Karrasch, 2007, Gnotobiotic IL-10−/−; NF- κBEGFP mice reveal the critical role of TLR/NF- κB signaling in commensal bacteria-induced colitis, J Immunol, 178, 6522, 10.4049/jimmunol.178.10.6522

Ko, 2007, Lactobacillus plantarum inhibits epithelial barrier dysfunction and interleukin-8 secretion induced by tumor necrosis factor- α, World J Gastroenterol, 13, 1962, 10.3748/wjg.v13.i13.1962

Lamine, 2004, Nitric oxide released by Lactobacillus farciminis improves TNBS-induced colitis in rats, Scand J Gastroenterol, 39, 37, 10.1080/00365520310007152

Lopez, 2008, Live and ultraviolet-inactivated Lactobacillus rhamnosus GG decrease flagellin-induced interleukin-8 production in Caco-2 cells, J Nutr, 138, 2264, 10.3945/jn.108.093658

Ma, 2004, TNF- α-induced increase in intestinal epithelial tight junction permeability requires NF- κB activation, Am J Physiol Gastrointest Liver Physiol, 286, G367, 10.1152/ajpgi.00173.2003

Marano, 1998, Tumor necrosis factor- α increases sodium and chloride conductance across the tight junction of CACO-2 BBE, a human intestinal epithelial cell line, J Membr Biol, 161, 263, 10.1007/s002329900333

McGuckin, 2009, Intestinal barrier dysfunction in inflammatory bowel diseases, Inflamm Bowel Dis, 15, 100, 10.1002/ibd.20539

Nenci, 2007, Epithelial NEMO links innate immunity to chronic intestinal inflammation, Nature, 446, 557, 10.1038/nature05698

Neurath, 1996, Local administration of antisense phosphorothioate oligonucleotides to the p65 subunit of NF- κB abrogates established experimental colitis in mice, Nat Med, 2, 998, 10.1038/nm0996-998

Peterson, 1992, Characterization of the enterocyte-like brush border cytoskeleton of the C2BBe clones of the human intestinal cell line, Caco-2, J Cell Sci, 102, 581, 10.1242/jcs.102.3.581

Petrof, 2009, Bacteria-free solution derived from Lactobacillus plantarum inhibits multiple NF- κB pathways and inhibits proteasome function, Inflamm Bowel Dis, 15, 1537, 10.1002/ibd.20930

Qin, 2009, L. plantarum prevents enteroinvasive Escherichia coli-induced tight junction proteins changes in intestinal epithelial cells, BMC Microbiol, 9, 63, 10.1186/1471-2180-9-63

Resta-Lenert, 2006, Probiotics and commensals reverse TNF- α- and IFN- γ-induced dysfunction in human intestinal epithelial cells, Gastroenterology, 130, 731, 10.1053/j.gastro.2005.12.015

Round, 2009, The gut microbiota shapes intestinal immune responses during health and disease, Nat Rev Immunol, 9, 313, 10.1038/nri2515

Seth, 2008, Probiotics ameliorate the hydrogen peroxide-induced epithelial barrier disruption by a PKC- and MAP kinase-dependent mechanism, Am J Physiol Gastrointest Liver Physiol, 294, G1060, 10.1152/ajpgi.00202.2007

Spehlmann, 2009, Nuclear factor- κB in intestinal protection and destruction, Curr Opin Gastroenterol, 25, 92, 10.1097/MOG.0b013e328324f857

Stadnyk, 2002, Intestinal epithelial cells as a source of inflammatory cytokines and chemokines, Can J Gastroenterol, 16, 241, 10.1155/2002/941087

Tanabe, 2008, Bifidobacterium infantis suppresses proinflammatory interleukin-17 production in murine splenocytes and dextran sodium sulfate-induced intestinal inflammation, Int J Mol Med, 22, 181

Tao, 2006, Soluble factors from Lactobacillus GG activate MAPKs and induce cytoprotective heat shock proteins in intestinal epithelial cells, Am J Physiol Cell Physiol, 290, C1018, 10.1152/ajpcell.00131.2005

Utech, 2005, Mechanism of IFN- γ-induced endocytosis of tight junction proteins: myosin II-dependent vacuolarization of the apical plasma membrane, Mol Biol Cell, 16, 5040, 10.1091/mbc.e05-03-0193

Vanderpool, 2008, Mechanisms of probiotic action: Implications for therapeutic applications in inflammatory bowel diseases, Inflamm Bowel Dis, 14, 1585, 10.1002/ibd.20525

Van Itallie, 2006, Claudins and epithelial paracellular transport, Annu Rev Physiol, 68, 403, 10.1146/annurev.physiol.68.040104.131404

Wang, 2005, Interferon- γ and tumor necrosis factor- α synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression, Am J Pathol, 166, 409, 10.1016/S0002-9440(10)62264-X

Wang, 2006, IFN- γ-induced TNFR2 expression is required for TNF-dependent intestinal epithelial barrier dysfunction, Gastroenterology, 131, 1153, 10.1053/j.gastro.2006.08.022

Yan, 2002, Probiotic bacterium prevents cytokine-induced apoptosis in intestinal epithelial cells, J Biol Chem, 277, 50959, 10.1074/jbc.M207050200

Yan, 2010, Probiotics: progress toward novel therapies for intestinal diseases, Curr Opin Gastroenterol, 26, 95, 10.1097/MOG.0b013e328335239a

Yan, 2007, Soluble proteins produced by probiotic bacteria regulate intestinal epithelial cell survival and growth, Gastroenterology, 132, 562, 10.1053/j.gastro.2006.11.022

Zareie, 2005, Novel effects of the prototype translocating Escherichia coli, strain C25 on intestinal epithelial structure and barrier function, Cell Microbiol, 7, 1782, 10.1111/j.1462-5822.2005.00595.x

Zhang, 2005, Alive and dead Lactobacillus rhamnosus GG decrease tumor necrosis factor- α-induced interleukin-8 production in Caco-2 cells, J Nutr, 135, 1752, 10.1093/jn/135.7.1752

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Microbiology (United Kingdom)

Tập 156 Số 11

3288-3297

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