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Research on The Effect of Intraepitherial Lymphocytes on Intestinal Mucosal Permeability

Research Project

Project/Area Number 12671278
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Digestive surgery
Research InstitutionOsaka University (2001-2002)
Kinki University (2000)

Principal Investigator

USUI Noriaki  Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助手 (30273626)

Co-Investigator(Kenkyū-buntansha) KUBOTA Akio  Osaka Medical Center and Research Institute for Maternal and Child Health, Department of Pediatric Surgery, Department Director, 部長(研究職) (10161671)
大柳 治正  近畿大学, 医学部, 教授 (00030958)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsMucosal Permeability / Gut Associated Lymphoid Tissue / Intestinal Mucosal Immunit / Intraepithelial Lymphocytes / Co-culture / Caco-2 / TEER / 腸管免疫 / CaCo-2 / co-culture
Research Abstract

Purpose : The aim of this study is to evaluate the influence of gut associated lymphoid tissue (GALT) on intestinal mucosal permeability by using a co-culture system of Caco-2 human enterocytes monolayer with lymphocytes harvested from normal and inflammatory SD rats
Materials and methods : Human Caco-2 enterocytes were seeded onto the lower aspect of porous membranes of a two-chamber system. "They were cultured for 10 14 days and allowed to form a polarized monolayer. After the measwement of transepithelial electric resistance (TEER) by using an epithelial voltohmmeter, Raji cells (a human lymphoid cell line with predominant B cell features), lymphocytes harvested from rat intestinal Peyer's patches (PPL), lamina propria lymphocytes (LPL) and intraepithelial lymphocytes (IEL) were added to the upper surface of the membranes and allowed to migrate through the pores in between the Caco-2 cell monolayers. After incorporation of Lymphocytes, TEERs were monitored for 3 days. PPL, LPL and IE … More L were also harvested from the bowel of indometacin induced inflammatory SD rats, in order to investigate the influence of bowel inflammation on these lymphocytes
Results : TEERs were elevated to 200-300 ohm.cm^2 14 days after seeding 1 x10^6 of Caco-2 cells per well, implying the integrity of tight junction. Co-culture with Raji cells for 3 days caused a decrease in TEERs, in a dose-dependent, manner. Permeability to dextran blue was 0.0t±0.0%, 0.0±0.0%, 0.0±0.0%, 0.6±0.3% and 15.411.5% at the Raji cell numbers of 0, 5x10^5 1x10^6, 2x10^6 and 4x10^6, respectively. Co-culture with 1x10^6 of PPL, 1x10^6 of LPL and 1x10^6 of IEL have not decreased TEERs of Caco-2 cell monolayeis. The intestine of SD rats that have been given Indometacin showed a thinner wall and fraglity, indicating an existence of bowel influnmation. Co-cultwe with PPL, LPL and IEL harvested from Indometacin induced inflammatory bowel also have not decreased TEERs of Caco-2 cell monolayers
Conclusion : GULT, such as PPL, LPL and IEL, deliverd from normal rats and inflammatory rats indicated no influence on the permeability of polarized human Caco-2 enterocyte monolayers. Further investigation may be required to clarify the mechanism of cell-cellular interaction between GALT and enterocytes Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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