Effetct of Pseudomonas aeruginosa-derived neutral ceramidase on immunological responses of three-dimensionally cultured human primary keratinocytes

• Project/Area Number: 24659293
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Pain science
• Research Institution: Juntendo University
• Principal Investigator: IWABUCHI Kazuhisa 順天堂大学, 公私立大学の部局等, 教授 (10184897)
• Co-Investigator(Kenkyū-buntansha): SUGA Yasushi 順天堂大学, 大学院医学研究科, 教授 (90245738)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: アトピー性皮膚炎 / 緑膿菌 / セラミダーゼ / ケラチノサイト / スフィンゴシン1リン酸 / 炎症性サイトカイン / TNFα / エンドセリン１ / 痒み / セラミド / 炎症 / 掻痒

Research Abstract

A Pseudomonas aeruginosa-derived neutral ceramidase (PaCDase) isolated from a patient with atopic dermatitis was shown to effectively degrade ceramide. The ceramide metabolite sphingosine-1-phosphate (S1P) stimulated the production of inflammatory mediators such as TNF and IL-8 from three-dimensionally cultured human primary keratinocytes (3D keratinocytes). PaCDase induced the production of TNF and IL-8, and this production was dependent on ceramidase enzymatic activity. S1P enhanced the gene expression of TNF and IL-8, and this enhancement was inhibited by a sphingosine kinase inhibitor and an S1P receptor antagonist. The TNF-binding antibody infliximab suppressed the PaCDase-induced upregulation of IL-8 mRNA but not of TNF mRNA. These findings suggest that, 3D keratinocytes produce S1P from sphingosine, which is produced through the hydrolysis of ceramide by PaCDase, and then S1P induced TNF production from these cells, resulted in production of IL-8.

Research Products

• [Journal Article] Pseudomonas-derived ceramidase induces production of inflammatory mediators from human keratinocytes via sphingosine-1-phosphate (2014)
  • Author(s): Oizumi A, Nakayama H, Okino N, Iwahara C, Kina K, Matsumoto R, Ogawa H, Takamori K, Ito M, Suga Y, Iwabuchi K.
  • Journal Title: PLoS One. Volume: 9(2) Issue: 2 Pages: 89402-89402
  • DOI: 10.1371/journal.pone.0089402
  • Peer Reviewed / Open Access
• [Journal Article] Organization and functions of glycolipid-enriched microdomains in phagocytes (2014)
  • Author(s): Ekyalongo R, Nakayama H, Kina K, Kaga N, Iwabuchi K.
  • Journal Title: BBA (Molecular and Cell Biology of Lipids) Volume: (in press)
• [Journal Article] Anti-neutral glycolipid antibodies in encephalomyeloradiculoneuropathy. (2014)
  • Author(s): Shima S, Kawamura N, Ishikawa T, Masuda H, Iwahara C, Niimi Y, Ueda A, Iwabuchi K, Mutoh T.
  • Journal Title: Neurology Volume: 82 Issue: 2 Pages: 114-118
  • DOI: 10.1212/wnl.0000000000000015
  • Peer Reviewed
• [Journal Article] Sevoflurane Suppresses TNF-α-Induced Inflammatory Responses in Small Airway Epithelial Cells under Anoxia/Reoxygenation Conditions. Bri (2013)
  • Author(s): W atanabe K. Iwahara C. Nakayama H. Iwabuchi K, Matsukawa T, Yokoyama K, Yamaguchi K, Kamiyama Y and Inada E.
  • Journal Title: J Anaesth Volume: 110(4) Pages: 637-45
• [Journal Article] GSL-enriched membrane microdomains in innate immune responses (2013)
  • Author(s): Nakayama H, Ogawa H, Takamori K, Iwabuchi K.
  • Journal Title: Arch. Immuno. et Therap. Exp Volume: 61(3) Pages: 217-28
• [Journal Article] Inhibition of connective tissue growth factor ameliorates rheumatoid arthritis in a murine model. (2013)
  • Author(s): Nozawa K, Fujishiro M, Kawasaki M, Yamaguchi A, Ikeda K, Morimoto S, Iwabuchi K, Yanagida M, Ichinose S, Morioka M, Ogawa H, Takamori K, Takasaki Y, Sekigawa I.
  • Journal Title: Arthritis Rheum. Volume: 65 Issue: 6 Pages: 1477-86
  • DOI: 10.1002/art.37902
  • Peer Reviewed
• [Journal Article] GSL-enriched membrane microdomains in innate immune responses. (2013)
  • Author(s): Nakayama H, Ogawa H, Takamori K, Iwabuchi K.
  • Journal Title: Arch Immunol Ther Exp (Warsz). Volume: 61 Issue: 3 Pages: 217-28
  • DOI: 10.1007/s00005-013-0221-6
  • Peer Reviewed
• [Journal Article] Sevoflurane suppresses tumour necrosis factor-α-induced inflammatory responses in small airway epithelial cells after anoxia/reoxygenation. (2013)
  • Author(s): Watanabe K, Iwahara C, Nakayama H, Iwabuchi K, Matsukawa T, Yokoyama K, Yamaguchi K, Kamiyama Y, Inada E.
  • Journal Title: Br J Anaesth Volume: 110 Issue: 4 Pages: 637-645
  • DOI: 10.1093/bja/aes469
  • Peer Reviewed
• [Journal Article] The novel neutrophil differentiation marker phosphatidylglucoside is involved in Fas-dependent apoptosis (2012)
  • Author(s): Kina K, Masuda H, Nakayama H, Iwahara C, Nagatsuka Y, Hirabayashi Y, Ogawa H, Takamori K, and Iwabuchi K.
  • Journal Title: Inflammation and Regeneration Volume: 32 Pages: 213-221
  • NAID: 130004482302
• [Journal Article] Role of stromal microenvironment in nonpharmacological resistance of CML to imatinib through Lyn/CXCR4 interactions in lipid rafts. (2012)
  • Author(s): Tabe Y, Jin L, Iwabuchi K, Wang RY, Ichikawa N, Miida T, Cortes J, Andreeff M, Konopleva M.
  • Journal Title: Leukemia Volume: 26 Issue: 5 Pages: 883-892
  • DOI: 10.1038/leu.2011.291
  • Peer Reviewed
• [Journal Article] Membrane microdomains in immunity: glycosphingolipid-enriched domain-mediated innate immune responses. (2012)
  • Author(s): Iwabuchi K, Nakayama H, Masuda H, Kina K, Ogawa H, Takamori K.
  • Journal Title: Biofactors Volume: 38 Issue: 4 Pages: 228-237
  • DOI: 10.1002/biof.1017
  • Peer Reviewed
• [Journal Article] Interplay of neuropilin-1 and semaphorin 3A after partial hepatectomy in rats. (2012)
  • Author(s): Fu L, Kitamura T, Iwabuchi K, Ichinose S, Yanagida M, Ogawa H, Watanabe S, Maruyama T, Suyama M, Takamori K.
  • Journal Title: World J Gastroenterol. Volume: 18 Issue: 36 Pages: 5034-41
  • DOI: 10.3748/wjg.v18.i36.5034
  • Peer Reviewed
• [Journal Article] Glycosphingolipid-Receptor Interactions in the Innate Immune Responses.
  • Author(s): Nakayama H, Iwabuchi K.
  • Journal Title: Glycoscience Biology and Medicine Volume: (in press)
• [Journal Article] Involvement of glycosphingolipid-enriched lipid rafts in inflammatory responses
  • Author(s): Iwabuchi K.
  • Journal Title: Front Biosci (Landmark Ed) Volume: (in press)
• [Presentation] Role of Glycolipids in Innate Immune Response. (2013)
  • Author(s): Iwabuchi K.
  • Organizer: 22nd International Symposium on Glycoconjugates
  • Place of Presentation: Dalian, China,
  • Invited
• [Presentation] Organization and functions of glycolipid-enriched microdomains in phagocytes. (2013)
  • Author(s): Iwabuchi K.
  • Organizer: 54th International Conference on the Bioscience of Lipids: Linking Transcription to Physiology in Lipidomics.
  • Place of Presentation: Bari, Italy
  • Invited
• [Presentation] 緑膿菌由来セラミダーゼによる三次元培養表皮シート・ケラチノサイトのサイトカイン産生機構について (2012)
  • Author(s): 泉亜美、岩原知博、喜納勝成、須賀康、沖野望、伊東信、小川秀興、髙森建二、岩渕和久
  • Organizer: 第54回日本脂質生化学会大会
  • Place of Presentation: 福岡
• [Presentation] スフィンゴシン一リン酸によるヒト表皮ケラチノサイトの遺伝子発現応答への影響について
  • Author(s): 岩渕和久、須賀康
  • Organizer: 医療看護研究会
  • Place of Presentation: 浦安市
• [Remarks]
  • URL: http://www.juntendo.ac.jp/graduate/nurs/research/profile_12.html