Synthesis of bacterial peptidoglycan fragments for elucidation of the immunostimulatory function and for aplication as immune adjuvant

• Project/Area Number: 17510178
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Living organism molecular science
• Research Institution: Osaka University
• Principal Investigator: FUJIMOTO Yukari Osaka University, Graduate School of Science, Department of Chemistry, Associate Professor, 大学院・理学研究科, 講師 (00362616)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,800,000 (Direct Cost: ¥3,800,000); Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: peptidoglycan / innate immunity / glycoconjugate / immune adjuvant / fluorescence-label

Research Abstract

Bacterial cell wall component, peptidoglycan (PGN), has been known as a potent immunostimulator and an immune adjuvant. In order to elucidate the recognition mechanism of PGN by its receptors such as Nod1, Nod2 and also Toll like receptor (TLR) 2, we synthesized the partial structures of PGN and the fluorescence-labeled compounds. The synthetic methods to construct the glycan-peptide structures were also developed.
For the investigation of Nod1 recognition, we synthesized the ligands, four optical isomers of γ-D-glutamyl-diaminopimelic acid (iE-DAP) using three optical isomers of DAP separated with chiral HPL C. The Nod1 stimulatory activities were obtained, and it was shown that the differences of the activities of four isomers. Based on this result, we developed the new synthetic method to obtain most stimulatory native meso-DAP and iE-meso-DAP. The fluorescence-labeled iE-DAP were also synthesized and succeeded in obtaining the labeled compound which kept the Nod1 stimulating activity. We also prepared the fluorescence-labeled Nod2 ligands, muramyl dipeptide (MDP), for the elucidation of Nod2 stimulation mechanism.
The PGN fragment, MDP has immune adjuvant activity, and we planed to apply its activity to a synthetic vaccine with making covalent connection between the adjuvant and antigen via a carrier protein. For the introduction of the immune' adjuvant to the complex, we developed the method to introduce the linker to the PGN fragments and succeded to obtain several immunostimulatory compounds including 6-O-acyl-MDP with the linker.

Research Products

• [Journal Article] A role of lipophilic peptidoglycan-related molecules in induction of Nodl-mediated immune responses (2007)
  • Author(s): M.Hasegawa
  • Journal Title: Journal of Biological Chemistry 282 Pages: 11757-11764
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A role of lipophilic peptidoglycan-related molecules in induction of Nod1-mediated immune responses. (2007)
  • Author(s): M.Hasegawa, A.Kawasaki, K.Yang, Y.Fujimoto, J.Masumoto, E.Breukink, G.Nufiez, K.Fukase, N.Inohara
  • Journal Title: J.Biol.Chem. 282 Pages: 11757-11764
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A role of lipophilic peptidoglycan-related molecules in induction of Nod1-mediated immune responses (2007)
  • Author(s): M.Hasegawa
  • Journal Title: Journal of Biological Chemistry (in press)
• [Journal Article] Differential release and distribution of Nod1 and Nod2 immunostimulatory molecules among bacterial species and environments (2006)
  • Author(s): M.Hasegawa
  • Journal Title: Journal of Biological Chemistry 281 Pages: 29054-29063
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Meso-Diaminopimelic Acid and meso-Lanthionine, Amino Acids Specific to Bacterial Peptidoglycans, Activates Human Epithelial Cells through NOD1 (2006)
  • Author(s): A.Uehara
  • Journal Title: J. Immunol. 177 Pages: 1796-1804
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Sarcoidosis and NOD1 variation with impaired recognition of intracellular Propionibacterium acnes. (2006)
  • Author(s): T.Tanabe
  • Journal Title: Biochim Biophys Acta 1762 Pages: 794-801
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Toll-like Receptors, NOD1 and NOD2, in Oral Epithelial Cells. (2006)
  • Author(s): Y.Sugawara
  • Journal Title: J. Dent. Res. 85 Pages: 524-529
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthesis of peptidoglycan fragments and evaluation of their biological activity. (2006)
  • Author(s): Seiichi Inamura
  • Journal Title: Organic ＆ Biomolecular Chemistry 4 Pages: 232-242
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A synthetic peptidoglycan fragment as a competitive inhibitor of the melanization cascade (2006)
  • Author(s): Ji W.Park
  • Journal Title: Journal of Biological Chemistry (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Nodl acts as an intracellular receptor to stimulate chemokine production and neutrophil recruitment in vivo. (2006)
  • Author(s): Junya Masumoto
  • Journal Title: The Journal of Experimental Medicine 203(1) Pages: 203-213
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthesis and bioactivity of fluorescence- and biotin-labeled lipid A analogues for investigation of recognition mechanism in innate immunity. (2006)
  • Author(s): Yukari Fujimoto
  • Journal Title: Tetrahedron Letters 47 Pages: 539-543
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthesis of peptidoglycan fragments and evaluation of their biological activity. (2006)
  • Author(s): Inamura, S., Fujimoto.Y., Kawasaki, A., Shiokawa, Z., Woelk, E., Heine, H., Lindner, B., Inohara, N., Kusumoto, S., Fukase, K.
  • Journal Title: Org.Biomol.Chem. 4 Pages: 232-242
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Differential release and distribution of Nod1 and Nod2 immunostimulatory molecules among bacterial species and environments. (2006)
  • Author(s): Hasegawa, M., Yang, K., Hashimoto, M., Park, J.-H., Kim, Y.-G., Fujimoto, Y., Nunez, G., Fukase, K., Inohara, N.
  • Journal Title: J.Biol.Chem. 281 Pages: 29054-29063
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Sarcoidosis and NOD1 variation with impaired recognition of intracellular Propionibacterium acnes. (2006)
  • Author(s): Tanabe, T., Ishige, I., Suzuki, Y., Aita, Y., Furukawa, A., Ishige, Y., Uchida, K., Suzuki, T., Takemura, T., Ikushima, S., Oritsu, M., Yokoyama, T., Fujimoto, Y., Fukase, K., Inohara, N., Nunez, G., Eishi, Y.
  • Journal Title: Biochim.Biophys.Acta 1762 Pages: 794-801
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Meso-diaminopimelic acid and meso-lanthionine, amino acids specific to bacterial peptidoglycans, activate human epithelial cells through NOD1. (2006)
  • Author(s): Uehara, A., Fujimoto.Y., Kawasaki, A., Kusumoto, S., Fukase, K., Takada, H.
  • Journal Title: J.Immunol. 177 Pages: 1796-1804
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Toll-like receptors, NOD1 and NOD2, in oral epithelial cells. (2006)
  • Author(s): Sugawara, Y., Uehara, A., Fujimoto.Y., Kusumoto, S., Fukase, K., Shibata, K., Sugawara, S., Sasano, T., Takada, H.
  • Journal Title: J.Dent.Res. 85 Pages: 524-529
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A synthetic peptidoglycan fragment as a competitive inhibitor of the melanization cascade. (2006)
  • Author(s): Park, J.W., Je, B-R., Piao, S., Inamura, S., Fujimoto, Y., Fukase, K,.Kusumoto, S., Soderhall, K., Ha, N.-C., Lee, B.L.
  • Journal Title: J.Biol.Chem. 281 Pages: 7747-7755
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nod1 acts as an intracellular receptor to stimulate chemokine production and neutrophil recruitment in vivo. (2006)
  • Author(s): Masumoto, J., Yang, K., Varambally, S., Hasegawa, M., Tomlins, S.A., Qui, S., Fujimoto.Y., Kawasaki, A., Foster, S.J., Horie, Y., Mak, T.W., Nunez, G., Chinnaiyan, A.M., Fukase, K., Inohara, N.
  • Journal Title: J.Exp.Med. 203 Pages: 203-213
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis and bioactivity of fluorescence-and biotin-labeled lipid A analogues for investigation of recognition mechanism in innate immunity. (2006)
  • Author(s): Fujimoto.Y., Kimura, E., Murata, S., Kusumoto, S., Fukase, K.
  • Journal Title: Tetrahedron Lett. 47 Pages: 539-543
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nod1 acts as an intracellular receptor to stimulate chemokine production and neutrophil recruitment in vivo. (2006)
  • Author(s): Junya Masumoto
  • Journal Title: The Journal of Experimental Medicine 203(1) Pages: 203-213
• [Journal Article] Synthesis of peptidoglycan fragments and evaluation of their biological activity. (2006)
  • Author(s): Seiichi Inamura
  • Journal Title: Organic & Biomolecular Chemistry 4 Pages: 232-242
• [Journal Article] Chemically synthesized pathogen-associated molecular patterns increase the expression of peptidoglycan recognition proteins viatoll-like receptors, NOD1 and NOD2 in human oral epithelial cells. (2005)
  • Author(s): Akiko Uehara
  • Journal Title: Cellular Microbiology 7 Pages: 675-686
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthesis of lipid A and its analogues for investigation of the structural basis for their bioactivity. (2005)
  • Author(s): Yukari Fujimoto
  • Journal Title: Journal of Endotoxin Research 11 Pages: 341-347
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human peptidoglycan recognition protein S is an effector of neutrophil-mediated innate immunity. (2005)
  • Author(s): Ju Hyun Cho
  • Journal Title: Blood 106 Pages: 2551-2558
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human peptidoglycan recognition protein S is an effector of neutrophil-mediated innate immunity. (2005)
  • Author(s): Cho, J.H., Fraser, I.P., Fukase, K., Kusumoto, S., Fujimoto, Y., Stahl, G.L., Ezekowitz, R.A.B.
  • Journal Title: Blood 106 Pages: 2551-2558
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Chemically synthesized pathogen-associated molecular patterns increase the expression of peptidoglycan recognition proteins via toll-like receptors, NOD1 and NOD2 in human oral epithelial cells. (2005)
  • Author(s): Akiko Uehara
  • Journal Title: Cellular Microbiology 7 Pages: 675-686
• [Book] エンドトキシン研究9「ヒト口腔上皮細胞のTLR系ならびにNOD系分子発現とその機能-特に抗菌物質産生について」 (2006)
  • Author(s): 上原亜希子
  • Total Pages: 6
  • Publisher: 医学図書出版株式会社(東京)
  • Description: 「研究成果報告書概要(和文)」より
• [Book] エンドトキシン研究9 「ヒトロ腔上皮細胞のTLR系ならびにNOD系分子発現とその機能-特に抗菌物質産生について」 (2006)
  • Author(s): 上原亜希子
  • Total Pages: 6
  • Publisher: 医学図書出版株式会社(東京)
• [Patent(Industrial Property Rights)] Modulator of Nodl Signaling (2005)
  • Inventor(s): 猪原直弘, 深瀬浩一, 藤本ゆかり
  • Industrial Property Rights Holder: Univ. of Michigan, 大阪大学
  • Filing Date: 2005-10-13
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] Modulator of Nod1 Signaling (2005)
  • Inventor(s): 猪原直弘, 深瀬浩一, 藤本ゆかり
  • Industrial Property Rights Holder: Univ. of Michigan 大阪大学
  • Filing Date: 2005-10-13