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Digestive Diseases and Sciences

, Volume 52, Issue 11, pp 2963–2971 | Cite as

Clostridium butyricum TO-A Culture Supernatant Downregulates TLR4 in Human Colonic Epithelial Cells

  • Atsushi Isono
  • Tatsuro Katsuno
  • Toru Sato
  • Tomoo Nakagawa
  • Yasutaka Kato
  • Naoki Sato
  • Gen’ichiro Seo
  • Yasuo Suzuki
  • Yasushi Saito
Original Paper

Abstract

The present study was performed to examine whether probiotics affect Toll-like receptor 4 (TLR4) expression in human colonic epithelial cells. Culture supernatants or heat-killed bacteria of Bacillus mesentericus TO-A, Clostridium butyricum TO-A, and Streptococcus faecalis T-110 were applied to human colonic epithelial cells. Treatment with C. butyricum TO-A culture supernatant significantly reduced TLR4 mRNA level (×0.16), even in the presence of interferon-γ (IFN-γ; ×0.21) as compared with untreated controls. High-performance liquid chromatography analysis showed that C. butyricum TO-A supernatant contains formate, acetate, and butyrate. Interestingly, TLR4 mRNA was significantly suppressed (×0.15–×0.22) only when cells were treated with solutions containing butyrate. Electrophoretic mobility shift assay suggested that the binding affinity of PU.1 to the promoter region of the TLR4 gene was markedly inhibited when the cells were treated with butyrate. This study suggested that butyrate produced by C. butyricum TO-A downregulates TLR4 mRNA level in human colonic epithelial cells.

Keywords

Toll-like receptor 4 HT-29 human colonic epithelial cells Clostridium butyricum TO-A Butyrate PU.1 

References

  1. 1.
    Aderem A, Ulevitch RJ (2000) Toll-like receptors in the induction of the innate immune response. Nature 406:782–787PubMedCrossRefGoogle Scholar
  2. 2.
    Takeda K, Kaisho T, Akira S (2003) Toll-like receptors. Annu Rev Immunol 21:335–376PubMedCrossRefGoogle Scholar
  3. 3.
    Akira S, Takeda K (2004) Toll-like receptor signalling. Nat Rev Immunol 4:499–511PubMedCrossRefGoogle Scholar
  4. 4.
    Medzhitov R (2001) Toll-like receptors and innate immunity. Nat Rev Immunol 1:135–145PubMedCrossRefGoogle Scholar
  5. 5.
    Akira S, Takeda K, Kaisho T (2001) Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol 2:675–680PubMedCrossRefGoogle Scholar
  6. 6.
    Banchereau J, Briere F, Caux C, Davoust J, Lebecque S, Liu YJ, Pulendran B, Palucka K (2000) Immunobiology of dendritic cells. Annu Rev Immunol 18:767–811PubMedCrossRefGoogle Scholar
  7. 7.
    Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392:245–252PubMedCrossRefGoogle Scholar
  8. 8.
    Cario E, Podolsky DK (2000) Differential alteration in intestinal epithelial cell expression of toll-like receptor 3 (TLR3) and TLR4 in inflammatory bowel disease. Infect Immun 68:7010–7017PubMedCrossRefGoogle Scholar
  9. 9.
    Ortega-Cava CF, Ishihara S, Rumi MA, Kawashima K, Ishimura N, Kazumori H, Udagawa J, Kadowaki Y, Kinoshita Y (2003) Strategic compartmentalization of Toll-like receptor 4 in the mouse gut. J Immunol 170:3977–3985PubMedGoogle Scholar
  10. 10.
    Boone DL, Ma A (2003) Connecting the dots from Toll-like receptors to innate immune cells and inflammatory bowel disease. J Clin Invest 111:1284–1286PubMedCrossRefGoogle Scholar
  11. 11.
    Kobayashi M, Kweon MN, Kuwata H, Schreiber RD, Kiyono H, Takeda K, Akira S (2003) Toll-like receptor-dependent production of IL-12p40 causes chronic enterocolitis in myeloid cell-specific Stat3-deficient mice. J Clin Invest 111:1297–1308PubMedCrossRefGoogle Scholar
  12. 12.
    Fedorak RN, Madsen KL (2004) Probiotics and the management of inflammatory bowel disease. Inflamm Bowel Dis 10:286–299PubMedCrossRefGoogle Scholar
  13. 13.
    Goossens D, Jonkers D, Stobberingh E, van den Bogaard A, Russel M, Stockbrugger R (2003) Probiotics in gastroenterology: indications and future perspectives. Scand J Gastroenterol Suppl:15–23Google Scholar
  14. 14.
    Kruis W (2004) Review article: antibiotics and probiotics in inflammatory bowel disease. Aliment Pharmacol Ther 20(Suppl 4):75–78PubMedCrossRefGoogle Scholar
  15. 15.
    Sartor RB (2004) Therapeutic manipulation of the enteric microflora in inflammatory bowel diseases: antibiotics, probiotics, and prebiotics. Gastroenterology 126:1620–1633PubMedCrossRefGoogle Scholar
  16. 16.
    Shanahan F (2005) Physiological basis for novel drug therapies used to treat the inflammatory bowel diseases I. Pathophysiological basis and prospects for probiotic therapy in inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol 288:G417–421PubMedCrossRefGoogle Scholar
  17. 17.
    Rembacken BJ, Snelling AM, Hawkey PM, Chalmers DM, Axon AT (1999) Non-pathogenic Escherichia coli versus mesalazine for the treatment of ulcerative colitis: a randomised trial. Lancet 354:635–639PubMedCrossRefGoogle Scholar
  18. 18.
    Gionchetti P, Rizzello F, Helwig U, Venturi A, Lammers KM, Brigidi P, Vitali B, Poggioli G, Miglioli M, Campieri M (2003) Prophylaxis of pouchitis onset with probiotic therapy: a double-blind, placebo-controlled trial. Gastroenterology 124:1202–1209PubMedCrossRefGoogle Scholar
  19. 19.
    Gionchetti P, Rizzello F, Venturi A, Brigidi P, Matteuzzi D, Bazzocchi G, Poggioli G, Miglioli M, Campieri M (2000) Oral bacteriotherapy as maintenance treatment in patients with chronic pouchitis: a double-blind, placebo-controlled trial. Gastroenterology 119:305–309PubMedCrossRefGoogle Scholar
  20. 20.
    Guslandi M, Giollo P, Testoni PA (2003) A pilot trial of Saccharomyces boulardii in ulcerative colitis. Eur J Gastroenterol Hepatol 15:697–698PubMedCrossRefGoogle Scholar
  21. 21.
    Fedorak RN, Madsen KL (2004) Probiotics and prebiotics in gastrointestinal disorders. Curr Opin Gastroenterol 20:146–155PubMedCrossRefGoogle Scholar
  22. 22.
    Furrie E, Macfarlane S, Thomson G, Macfarlane GT (2005) Toll-like receptors-2, -3 and -4 expression patterns on human colon and their regulation by mucosal-associated bacteria. Immunology 115:565–574PubMedCrossRefGoogle Scholar
  23. 23.
    Maaser C, Heidemann J, von Eiff C, Lugering A, Spahn TW, Binion DG, Domschke W, Lugering N, Kucharzik T (2004) Human intestinal microvascular endothelial cells express Toll-like receptor 5: a binding partner for bacterial flagellin. J Immunol 172:5056–5062PubMedGoogle Scholar
  24. 24.
    Pedersen G, Andresen L, Matthiessen MW, Rask-Madsen J, Brynskov J (2005) Expression of Toll-like receptor 9 and response to bacterial CpG oligodeoxynucleotides in human intestinal epithelium. Clin Exp Immunol 141:298–306PubMedCrossRefGoogle Scholar
  25. 25.
    Lin M, Rikihisa Y (2003) Ehrlichia chaffeensis and Anaplasma phagocytophilum lack genes for lipid A biosynthesis and incorporate cholesterol for their survival. Infect Immun 71:5324–5331PubMedCrossRefGoogle Scholar
  26. 26.
    Lin M, Rikihisa Y (2004) Ehrlichia chaffeensis downregulates surface Toll-like receptors 2/4, CD14 and transcription factors PU.1 and inhibits lipopolysaccharide activation of NF-kappa B, ERK 1/2 and p38 MAPK in host monocytes. Cell Microbiol 6:175–186PubMedCrossRefGoogle Scholar
  27. 27.
    Abreu MT, Arnold ET, Thomas LS, Gonsky R, Zhou Y, Hu B, Arditi M (2002) TLR4 and MD-2 expression is regulated by immune-mediated signals in human intestinal epithelial cells. J Biol Chem 277:20431–20437PubMedCrossRefGoogle Scholar
  28. 28.
    Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, Miyake K, Kimoto M (1999) MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med 189:1777–1782PubMedCrossRefGoogle Scholar
  29. 29.
    Mita Y, Dobashi K, Endou K, Kawata T, Shimizu Y, Nakazawa T, Mori M (2002) Toll-like receptor 4 surface expression on human monocytes and B cells is modulated by IL-2 and IL-4. Immunol Lett 81:71–75PubMedCrossRefGoogle Scholar
  30. 30.
    Bosisio D, Polentarutti N, Sironi M, Bernasconi S, Miyake K, Webb GR, Martin MU, Mantovani A, Muzio M (2002) Stimulation of toll-like receptor 4 expression in human mononuclear phagocytes by interferon-gamma: a molecular basis for priming and synergism with bacterial lipopolysaccharide. Blood 99:3427–3431PubMedCrossRefGoogle Scholar
  31. 31.
    Wang T, Lafuse WP, Zwilling BS (2000) Regulation of toll-like receptor 2 expression by macrophages following Mycobacterium avium infection. J Immunol 165:6308–6313PubMedGoogle Scholar
  32. 32.
    Böcker U, Yezerskyy O, Feick P, Manigold T, Panja A, Kalina U, Herweck F, Rossol S, Singer MV (2003) Responsiveness of intestinal epithelial cell lines to lipopolysaccharide is correlated with Toll-like receptor 4 but not Toll-like receptor 2 or CD14 expression. Int J Colorectal Dis 18:25–32PubMedCrossRefGoogle Scholar
  33. 33.
    Laribee RN, Klemsz MJ (2001) Loss of PU.1 expression following inhibition of histone deacetylases. J Immunol 167:5160–5166PubMedGoogle Scholar
  34. 34.
    Rehli M, Poltorak A, Schwarzfischer L, Krause SW, Andreesen R, Beutler B (2000) PU.1 and interferon consensus sequence-binding protein regulate the myeloid expression of the human Toll-like receptor 4 gene. J Biol Chem 275:9773–9781PubMedCrossRefGoogle Scholar
  35. 35.
    Roger T, David J, Glauser MP, Calandra T (2001) MIF regulates innate immune responses through modulation of Toll-like receptor 4. Nature 414:920–924PubMedCrossRefGoogle Scholar
  36. 36.
    Inan MS, Rasoulpour RJ, Yin L, Hubbard AK, Rosenberg DW, Giardina C (2000) The luminal short-chain fatty acid butyrate modulates NF-kappaB activity in a human colonic epithelial cell line. Gastroenterology 118:724–734PubMedCrossRefGoogle Scholar
  37. 37.
    Luhrs H, Gerke T, Schauber J, Dusel G, Melcher R, Scheppach W, Menzel T (2001) Cytokine-activated degradation of inhibitory kappaB protein alpha is inhibited by the short-chain fatty acid butyrate. Int J Colorectal Dis 16:195–201PubMedCrossRefGoogle Scholar
  38. 38.
    Segain JP, Raingeard de la Bletiere D, Bourreille A, Leray V, Gervois N, Rosales C, Ferrier L, Bonnet C, Blottiere HM, Galmiche JP (2000) Butyrate inhibits inflammatory responses through NFkappaB inhibition: implications for Crohn's disease. Gut 47:397–403PubMedCrossRefGoogle Scholar
  39. 39.
    Yin L, Laevsky G, Giardina C (2001) Butyrate suppression of colonocyte NF-kappa B activation and cellular proteasome activity. J Biol Chem 276:44641–44646PubMedCrossRefGoogle Scholar
  40. 40.
    Luhrs H, Gerke T, Muller JG, Melcher R, Schauber J, Boxberge F, Scheppach W, Menzel T (2002) Butyrate inhibits NF-kappaB activation in lamina propria macrophages of patients with ulcerative colitis. Scand J Gastroenterol 37:458–466PubMedCrossRefGoogle Scholar
  41. 41.
    Scheppach W (1996) Treatment of distal ulcerative colitis with short-chain fatty acid enemas. A placebo-controlled trial. German-Austrian SCFA Study Group. Dig Dis Sci 41:2254–2259PubMedCrossRefGoogle Scholar
  42. 42.
    Scheppach W, Sommer H, Kirchner T, Paganelli GM, Bartram P, Christl S, Richter F, Dusel G, Kasper H (1992) Effect of butyrate enemas on the colonic mucosa in distal ulcerative colitis. Gastroenterology 103:51–56PubMedGoogle Scholar
  43. 43.
    Vernia P, Marcheggiano A, Caprilli R, Frieri G, Corrao G, Valpiani D, Di Paolo MC, Paoluzi P, Torsoli A (1995) Short-chain fatty acid topical treatment in distal ulcerative colitis. Aliment Pharmacol Ther 9:309–313PubMedCrossRefGoogle Scholar
  44. 44.
    Vernia P, Monteleone G, Grandinetti G, Villotti G, Di Giulio E, Frieri G, Marcheggiano A, Pallone F, Caprilli R, Torsoli A (2000) Combined oral sodium butyrate and mesalazine treatment compared to oral mesalazine alone in ulcerative colitis: randomized, double-blind, placebo-controlled pilot study. Dig Dis Sci 45:976–981PubMedCrossRefGoogle Scholar
  45. 45.
    Okamoto T, Sasaki M, Tsujikawa T, Fujiyama Y, Bamba T, Kusunoki M (2000) Preventive efficacy of butyrate enemas and oral administration of Clostridium butyricum M588 in dextran sodium sulfate-induced colitis in rats. J Gastroenterol 35:341–346PubMedCrossRefGoogle Scholar
  46. 46.
    Katakura K, Lee J, Rachmilewitz D, Li G, Eckmann L, Raz E (2005) Toll-like receptor 9-induced type I IFN protects mice from experimental colitis. J Clin Invest 115:695–702PubMedCrossRefGoogle Scholar
  47. 47.
    Rachmilewitz D, Katakura K, Karmeli F, Hayashi T, Reinus C, Rudensky B, Akira S, Takeda K, Lee J, Takabayashi K, Raz E (2004) Toll-like receptor 9 signaling mediates the anti-inflammatory effects of probiotics in murine experimental colitis. Gastroenterology 126:520–528PubMedCrossRefGoogle Scholar
  48. 48.
    Rakoff-Nahoum S, Paglino J, Eslami-Varzaneh F, Edberg S, Medzhitov R (2004) Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell 118:229–241PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2006

Authors and Affiliations

  • Atsushi Isono
    • 1
  • Tatsuro Katsuno
    • 1
  • Toru Sato
    • 1
  • Tomoo Nakagawa
    • 1
  • Yasutaka Kato
    • 1
  • Naoki Sato
    • 2
  • Gen’ichiro Seo
    • 2
  • Yasuo Suzuki
    • 3
  • Yasushi Saito
    • 1
  1. 1.Department of Clinical Cell Biology (F5)Graduate School of Medicine, Chiba UniversityChibaJapan
  2. 2.Toa Pharmaceutical Co., Ltd.TatebayashiJapan
  3. 3.Department of Internal MedicineToho University Sakura Medical CenterChibaJapan

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