Research on The Effect of Intraepitherial Lymphocytes on Intestinal Mucosal Permeability

上皮内淋巴细胞对肠粘膜通透性影响的研究

• 批准号: 12671278
• 负责人: USUI Noriaki
• 金额: $ 1.86万
• 依托单位: Osaka University (2001-2002)Kinki University (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671278/
• 关键词: Mucosal Permeability; Gut Associated Lymphoid Tissue; Intestinal Mucosal Immunit; Intraepithelial Lymphocytes; Co-culture; Caco-2; TEER; 腸管免疫; CaCo-2; co-culture

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 ratsMaterials 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 lymphocytesResults : 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 monolayersConclusion : 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

目的：利用Caco-2人肠上皮细胞单层与正常及炎症性SD大鼠淋巴细胞共培养体系，观察肠道相关淋巴组织(GALT)对肠粘膜通透性的影响。材料与方法：将人Caco-2肠上皮细胞接种于两室多孔膜下部。他们被培养了10-14天，并被允许形成一个偏振单分子层。用上皮伏安计测量跨皮电阻(TEER)后，将Raji细胞(一种以B细胞为主的人淋巴样细胞系)、大鼠肠道Peyer‘s结(PPL)、固有层淋巴细胞(LPL)和上皮内淋巴细胞(IEL)加入到膜的上表面，并允许它们通过Caco-2细胞单层之间的孔迁移。淋巴细胞掺入后，对受试者监测3天。Ppl、lpl和IE…结果：接种1×10~(-6)个/孔的Caco-2细胞14天后，TERS升高到200~300欧姆·cm~(-2)，提示紧密连接的完整性。与Raji细胞共培养3天后，TERs以剂量依赖的方式减少。Raji细胞数为0、5×10~(-5)、1×10~(-6)、2×10~(-6)和4×10~(-6)时，对葡聚糖蓝的通透性分别为0.0t±0.0%、0.0±0.0%、0.0±0.0%、0.6±0.3%和15.411.5%。1×10~(-6)的PPL、1×10~(-6)的LPL和1×10~(-6)的IEL共培养不能降低Caco-2细胞的单层数。给予吲哚美辛的SD大鼠的肠壁较薄，肠壁较脆，提示肠炎的存在。结论：正常大鼠和炎症大鼠分泌的PPL、LPL和IEL对人Caco-2细胞单层通透性无明显影响。可能需要进一步的研究来澄清GALT和肠细胞之间的细胞间相互作用的机制