Molecular analyzes of sensitization phase of contact hypersensitivity

接触性超敏反应致敏期的分子分析

基本信息

批准号：
13672273
负责人：
HIGASHI Nobuaki
金额：
$ 2.62万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672273/
关键词：
macrophage antigen presentation lymph node contact hypersensitivity sialoadhesin lectin cell trafficking dendritic cells 糖鎖感作 COX-2

项目摘要

The aim of this proposal was to elucidate molecular mechanisms of sensitization phase of contact sensitivity, especially focusing on the entry of dermal macrophages to the draining lymph node and on the biological function of the cells inside the lymph node during the sensitization phase.(1) Entry of dermal macrophages to the draining lymph node : Involvement of lectin-carbohydrate interaction in the entry of macrophages expressing macrophage galactose-type C-type lectin (MGL1) in the sensitization phase was investigated. Immunohistochemical localization of binding sites of recombinant MGL1 in LNs was revealed to be similar to distribution of cells expressing MGL1 apparently migrated from the dermis during sensitization. Affinity chromatography with immobilized recombinant MGL1 successfully purified a predominant component that was identified as sialoadhesin. Immobilize sialoadhesin actually bound recombinant MGL1 as well as MGL1-transfected CHO cells. The binding was dependent on the carbohydrates expressed on sialoadhesin. Immunohistochemical localization of sialoadhesin and MGL1 ligand coincided. Furthermore, sialoadhesin and MGL1 closely distributed in subcapsular sinus.(2) Biological function of the dermal macrophages inside the lymph node during the sensitization phase : The aim of the study is to investigate how the MGL1 + cells modulate the sensitization phase. MGL+ cells were tested for the expression of cyclooxygenase (COXJ-2 mRNA. The cells expressed COX-2 mRNA and released PGE2, a final product of the COX-2-dependent prostanoids. The administration of a COX-2 inhibitor NS-398 during the sensitization phase inhibited the production of PGE2 in the lymph node and also antigen-dependent ear swelling of the sensitizad mice.

该提案的目的是阐明接触敏感性致敏阶段的分子机制，特别关注真皮巨噬细胞进入引流淋巴结以及致敏阶段淋巴结内细胞的生物学功能。(1)真皮巨噬细胞进入引流淋巴结：凝集素-碳水化合物相互作用参与表达巨噬细胞半乳糖型C型凝集素的巨噬细胞的进入研究了敏化阶段的（MGL1）。 LN 中重组 MGL1 结合位点的免疫组织化学定位与致敏期间明显从真皮迁移的表达 MGL1 的细胞的分布相似。使用固定化重组 MGL1 的亲和层析成功纯化了主要成分，该成分被鉴定为唾液酸粘附素。固定实际结合重组 MGL1 以及 MGL1 转染的 CHO 细胞的唾液酸粘附素。结合取决于唾液酸粘附素上表达的碳水化合物。唾液酸粘附素和 MGL1 配体的免疫组织化学定位一致。此外，唾液酸粘附素和MGL1紧密分布在囊下窦中。(2)致敏期淋巴结内真皮巨噬细胞的生物学功能：本研究的目的是研究MGL1 + 细胞如何调节致敏期。测试了 MGL+ 细胞的环加氧酶 (COXJ-2 mRNA) 的表达。细胞表达 COX-2 mRNA 并释放 PGE2（COX-2 依赖性前列腺素的最终产物）。在治疗期间给予 COX-2 抑制剂 NS-398致敏阶段抑制了淋巴结中 PGE2 的产生，也抑制了致敏小鼠的抗原依赖性耳肿胀。