Expression cloning of the gene that transform Caco-2 cells to M cells

• Project/Area Number: 22790343
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Human pathology
• Research Institution: Shinshu University
• Principal Investigator: KOBAYASHI Motohiro 信州大学, 医学系研究科, 講師 (00362137)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: M細胞 / 発現クローニング

Research Abstract

Our research goal was to clone the gene that transforms Caco-2 cells into M cells, which transcytose antigenic molecules in the intestinal lumen to subepithelial immune cells, by employing the Caco-2/Raji cells co-culture system. While we tried to find optimal condition for the induction of M cell phenotype, reproducible condition was not identified. Moreover, the transfection efficiency of Caco-2 cells by a lipofection method was too low to perform expression cloning.

Research Products

• [Journal Article] Reduced glycosylation ofα-dystroglycans on carcinoma cells contributes to formation of highly infiltrative histological patterns in prostate cancer (2011)
  • Author(s): Shimojo H, Kobayashi M, Kamigaito T, Shimojo Y, Fukuda M, Nakayama J
  • Journal Title: Prostate Volume: 71 Issue: 11 Pages: 1151-1157
  • DOI: 10.1002/pros.21330
  • Peer Reviewed
• [Journal Article] O-Glycans carrying non-sulfated sialyl Lewis X appearing on HEV-like vessels in gastric MALT lymphoma function as L-selectin ligands (2011)
  • Author(s): Kobayashi M, Mitoma J, Hoshino H, Yu SY, Shimojo Y, Suzawa K, Khoo KH, Fukuda M, Nakayama J
  • Journal Title: J Pathol Volume: 224 Issue: 1 Pages: 67-77
  • DOI: 10.1002/path.2851
  • Peer Reviewed
• [Journal Article] An integrinα4β7・IgG heterodimeric chimera binds to MAdCAM-1 on high endothelial venules in gut-associated lymphoid tissue (2011)
  • Author(s): Hoshino H, Kobayashi M, Mitoma J, Sato Y, Fukuda M, Nakayama J
  • Journal Title: J Histochem Cytochem Volume: 59 Issue: 6 Pages: 572-583
  • DOI: 10.1369/0022155411404416
  • Peer Reviewed
• [Journal Article] Membrane-associated activation of cholesterolα-glucosyltransferase, an enzyme responsible for biosynthesis of cholesteryl-α-D-glucopyranoside in Helicobacter pylori critical for its survival (2011)
  • Author(s): Hoshino H, Tsuchida A, Kametani K, Mori M, Nishizawa T, Suzuki T, Nakamura H, Lee H, Ito Y, Kobayashi M, Masumoto J, Fujita M, Fukuda M, Nakayama J
  • Journal Title: J Histochem Cytochem Volume: 59 Issue: 1 Pages: 98-105
  • DOI: 10.1369/jhc.2010.957092
  • Peer Reviewed
• [Journal Article] Induction of high endothelial venule-like vessels expressing GlcNAc6ST-1-mediated L-selectin ligand carbohydrate and mucosal addressin cell adhesion molecule 1(MAdCAM-1) in a mouse model of" Candidatus Helicobacter heilmannii"-induced gastritis and gastric mucosa-associated lymphoid tissue(MALT) lymphoma (2010)
  • Author(s): Suzuki A, Kobayashi M, Matsuda K, Matsumoto T, Kawakubo M, Kumazawa S, Koide N, Miyagawa S, Ota H
  • Journal Title: Helicobacter Volume: 15 Issue: 6 Pages: 538-548
  • DOI: 10.1111/j.1523-5378.2010.00801.x
  • Peer Reviewed
• [Journal Article] Novel anti-carbohydrate antibodies reveal the cooperative function of sulfated N-and O-glycans in lymphocyte homing (2010)
  • Author(s): Hirakawa J, Tsuboi K, Sato K, Kobayashi M, Watanabe S, Takakura A, Imai Y, Ito Y, Fukuda M, and Kawashima H.
  • Journal Title: J. Biol. Chem Volume: 285 Issue: 52 Pages: 40864-40878
  • DOI: 10.1074/jbc.m110.167296
  • Peer Reviewed
• [Journal Article] Immunohistochemical analysis of carbohydrate antigens in chronic inflammatory gastrointestinal diseases (2010)
  • Author(s): Kobayashi M(corresponding author), Nakayama J
  • Journal Title: Methods Enzymol Volume: 479 Pages: 271-289
  • DOI: 10.1016/s0076-6879(10)79016-9
  • ISBN: 9780123809971
  • Peer Reviewed
• [Journal Article] Induction of HEV-like vessels expressing GlcNAc6ST-1-mediated L-selectin ligand carbohydrate and MAdCAM-1 in a mouse model of "Candidatus Helicobacter heilmannii"-induced gastritis and MALT lymphoma (2010)
  • Author(s): Suzuki A, et al
  • Journal Title: Helicobacter Volume: 15 Pages: 538-548
  • Peer Reviewed
• [Journal Article] Novel anti-carbohydrate antibodies reveal the cooperative function of sulfated N- and O-glycans in lymphocyte homing (2010)
  • Author(s): Hirakawa J, et al
  • Journal Title: J Biol Chem Volume: 285 Pages: 40864-40878
  • Peer Reviewed
• [Presentation] Production of an antibody against long-form N-acetylglucosamine-6-ο-sulfotransfe rase 1 that is expressed in human high endothelial venules (2011)
  • Author(s): Fujiwara M, Kobayashi M, Hoshino H, Uchimura K, Sakai Y, Nakada T, Fukuda M, Nakayama J
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: Seattle, WA, USA
  • Year and Date: 2011-11-11
• [Presentation] Production of an antibody against long-form N-acetylglucosamine-6-O-sulfotransferase 1 that is expressed in human high endothelial venules (2011)
  • Author(s): Fujiwara M, et al
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: Seattle, WA, USA
  • Year and Date: 2011-11-11
• [Presentation] Prominent expression of sLeX-capped coreο-glycans on HEV-like vessels in gastric MALT lymphoma (2011)
  • Author(s): 小林基弘,中山淳
  • Organizer: 第100回日本病理学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2011-04-29
• [Presentation] Prominent expression of sLeX-capped core 2 O-glycans on HEV-like vessels in gastric MALT lymphoma (2011)
  • Author(s): 小林基弘, 他
  • Organizer: 第100回日本病理学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2011-04-29
• [Presentation] Cholesterylα-glucosides of Helicobacter pylori play critical roles in bacterial growth and innate immunity by invariant natural killer T cells (2010)
  • Author(s): Ito Y, Vela JL, Matumura F, Hoshino H, Tyznik A, Lee H, Kobayashi M, Seeberger P, Nakayama J, Kronenberg M, Fukuda M
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: St. Petersburg, FL, USA
  • Year and Date: 2010-11-10
• [Presentation] ο-glycans carrying non-sulfated sialyl Lewis X appearing on HEV-like vessels in gastric MALT lymphoma function as L-selectin ligands (2010)
  • Author(s): Kobayashi M, Mitoma J, Hoshino H, Yu SY, Shimojo Y, Suzawa K, Khoo KH, Fukuda M, Nakayama J
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: Petersburg, FL, USA
  • Year and Date: 2010-11-09
• [Presentation] Reduced glycosylation ofα-dystroglycans on carcinoma cells contributes to formation of highly infiltrative histological patterns in prostate cancer (2010)
  • Author(s): Shimojo H, Kobayashi M, Fukuda M, Nakayama J
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: St. Petersburg, FL, USA
  • Year and Date: 2010-11-09
• [Presentation] Deficiency of L-selectin ligand biosynthesis by knocking out of GlcNAc6ST-1 and-2 suppresses asthmatic reponse in murine model of asthma (2010)
  • Author(s): Bao X, Nakayama J, Miler M, Pham A, Kobayashi M, Kawashima H, Broide D, Fukuda M
  • Organizer: Annual Conference of the Society for Glycobiology
  • Place of Presentation: St. Petersburg, FL, USA
  • Year and Date: 2010-11-09
• [Presentation] O-glycans carrying non-sulfated sialyl Lewis X appearing on HEV-like vessels in gastric MALT lymphoma function as L-selectin ligands (2010)
  • Author(s): Kobayashi M, et al
  • Organizer: Glycobiology 2010
  • Place of Presentation: St.Petersburg, FL, USA
  • Year and Date: 2010-11-09