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FOOD ALLERGEN TRANSPORT BY HUMAN INTESTINAL CELLS : A STUDY USING CULTURED CACO-2 CELLS

人类肠道细胞的食物过敏原转运：使用培养的 CACO-2 细胞的研究

基本信息

批准号：
11680155
负责人：
MINAMI Hisanori
金额：
$ 2.5万
依托单位：
FACULTY OF ENVIRONMENTAL AND SYMBIOTIC SCIENCES, PREFECTURAL UNIVERSITY OF KUMAMOTO (2000-2001)Osaka Prefectural College of Nursing (1999)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

Besides functioning as a mucosal barrier and transporting nutrients, intestinal epithelial cells (IECs) also serve as antigen presenting cells (APCs). Modification of protein antigens by proteolysis is one of the principal steps in antigen presentation to Th cells. We used a Caco-2 intestinal epithelial cell line to investigate the transepithelial transport of the dietary antigen, ovalbumin (OVA). We also examined in Caco-2 cell layers, the effects of the proinflammatory cytokine interferon-γ (IFN-γ) on the antigen transport process. Caco-2 cell layers transferred both intact and degraded OVA from the mucosal to the serosal side. IFN-γ stimulated OVA transport and most of the transported OVA in such cells was degraded. We also examined OVA uptake by Caco-2 cells using immunohistochemical means. Caco-2 cells incorporated OVA in a time-dependent manner and IFN-γ significantly enhanced antigen internalization. Flow cytometry also demonstrated that IFN-γ elevated the internalization of FITC-OVA. We also determined the effect of low and high concentrations of IFN-γ on mucosal permeability and internalization of FITC-OVA. Although both 10 and 50 ng/mL IFN-γ stimulated mucosal permeability to same extent, more FITC-OVA was internalized by Caco-2 cells incubated with 50, than with 10 ng/mL IFN-γ. These results suggest that the effects of IFN-γ on mucosal permeability and on the internalization of antigens by intestinal epithelial cells are brought about by different mechanisms. Therefore, higher concentrations of IFN-γ stimulates the uptake, processing and transport of dietary antigens by IECs.

除了充当粘膜屏障和运输营养物质外，肠上皮细胞（IEC）还充当抗原呈递细胞（APC）。通过蛋白水解修饰蛋白质抗原是将抗原呈递给 Th 细胞的主要步骤之一。我们使用 Caco-2 肠上皮细胞系来研究饮食抗原卵清蛋白 (OVA) 的跨上皮转运。我们还在 Caco-2 细胞层中检查了促炎细胞因子干扰素-γ (IFN-γ) 对抗原转运过程的影响。 Caco-2 细胞层将完整的和降解的 OVA 从粘膜转移到浆膜侧。 IFN-γ 刺激 OVA 转运，并且此类细胞中大部分转运的 OVA 被降解。我们还使用免疫组织化学方法检查了 Caco-2 细胞对 OVA 的摄取。 Caco-2 细胞以时间依赖性方式掺入 OVA，并且 IFN-γ 显着增强抗原内化。流式细胞术还表明 IFN-γ 增强了 FITC-OVA 的内化。我们还确定了低浓度和高浓度的 IFN-γ 对粘膜通透性和 FITC-OVA 内化的影响。虽然 10 ng/mL 和 50 ng/mL IFN-γ 刺激粘膜通透性的程度相同，但与 10 ng/mL IFN-γ 相比，与 50 ng/mL IFN-γ 一起孵育的 Caco-2 细胞内化了更多的 FITC-OVA。这些结果表明，IFN-γ对粘膜通透性和肠上皮细胞抗原内化的影响是由不同的机制引起的。因此，较高浓度的 IFN-γ 会刺激 IEC 对饮食抗原的摄取、加工和运输。