Basic research on ex vivo modification of dendritic cell functions for development of treatment for inflammatory bowel diseases

树突状细胞功能的离体修饰的基础研究用于开发炎症性肠病的治疗

• 批准号: 13470124
• 负责人: DOHI Taeko
• 金额: $ 8.64万
• 依托单位: Research Institute, International Medical Center of Japan
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470124/
• 关键词: Ulcerative colitis; Dendritic cells; Peyer's patches; Cytokines; Chemokines; Commensal flora; lipopolysaccharides; inflammatory bowel disease; パイエル板

The purpose of this project was to clarify the function of dendritic cells (DC) in normal murine gastrointestinal (GI) tract as well as human GI tract in terms of the characteristic hyporesponsiveness to normal commensal flora, to develop a new remedy for inflammatory bowel disease.1.In a mouse model for ulcerative colitis (UC), recruitment of dendritic cells contributed to the perpetuation of disease, which was efficiently blocked by a soluble form chimeric protein of lymphotoxin β receptor.2.Human colonic macrophages and dendritic cells showed low cytokine responses after. stimulation with lipopolysaccharides (LPS), which was due to lack of MD-2, an essential molecule for LPS-signaling via toll like receptor 4. Further analysis of colonic cell obtained from UC patients showed aberrant expression of MD-2 and cytokines.3.We found that adoptively transferred DCs were localized in mesenteric lymph nodes. It indicated the possibility of modulation of mucosal immunity by exogenous DC.4.Because the significant side effect of retroviral vector was revealed during the research period, we searched other methods to modify function of DC ex vivo. We analyzed the effect of cholera toxin (CT) and CT was able to fully activate DC by activation NF-κB which was dependent on the binding to cell surface GM1 ganglioside.5.To develop a methods to deliver molecules into DC utilizing intracelluer transport, we created new double mCTs (dmCTs) that have two amino acid substitutions in the ADP-ribosyltransferase active center (E112K) and COOH-terminal KDEL (E112K/KDEV or E112K/KDGL). KDEL mutant was transported to Golgi but not to ER.Results of these serial experiment listed above provided important and basic knowledge to develop new remedies for the treatment of inflammatory bowel disease by modifying the function of dendritic cells ex vivo.

本课题旨在阐明树突状细胞（DC）在正常小鼠胃肠道和人胃肠道中对正常肠道植物群的低反应性的功能，以开发治疗炎症性肠病的新药物。1.在小鼠溃疡性结肠炎（UC）模型中，DC的募集有助于疾病的持续，该作用可被光敏素β受体可溶性嵌合蛋白有效阻断。用脂多糖（LPS）刺激，这是由于缺乏MD-2，MD-2是通过Toll样受体4进行LPS信号传导的必需分子。进一步分析UC患者结肠细胞MD-2和细胞因子的异常表达。3.过继转移的DC定位于肠系膜淋巴结。4.由于逆转录病毒载体在研究过程中暴露出明显的毒副作用，因此我们寻找其他方法来体外修饰DC的功能。我们分析了霍乱毒素（CT）对DC的作用，发现CT能够通过与细胞表面神经节苷脂GM 1结合激活NF-κB而完全激活DC。我们创建了新的双mCT（dmCT），其在ADP-核糖基转移酶活性中心（E112 K）和COOH-末端KDEL中具有两个氨基酸取代（E112K/KDEV或E112K/KDGL）。上述一系列实验结果为通过体外修饰树突状细胞的功能来开发治疗炎症性肠病的新药物提供了重要的基础知识。

项目成果

Lipopolysaccharide induces CD25-positive, IL-10-producing lymphocytes without secretion of proinflammatory cytokines in the human colon : Low MD-2 mRNA expression in the colonic macrophages.

脂多糖在人结肠中诱导 CD25 阳性、产生 IL-10 的淋巴细胞，而不分泌促炎细胞因子：结肠巨噬细胞中 MD-2 mRNA 表达低。

• 发表时间: 2004
• 期刊: J.Clin.Immunol. 24
• 作者: Shirasawa S;T;Shirai Y
• 通讯作者: Shirai Y

Elimination of colonic patches with lymphotoxin b receptor-Ig prevents Th2 cell-type colitis

用淋巴毒素 B 受体 Ig 消除结肠斑可预防 Th2 细胞型结肠炎

• 发表时间: 2001
• 期刊: J.Immunol. 167
• 作者: Shirasawa S;T;Shirai Y;Kawamura T;Dohi T;Hirotomo Kato;Yuki I.Kawamura;Taeko Dohi;Kato H;Kawamura YI;Dohi T;Hashimoto M;Taeko Dohi
• 通讯作者: Taeko Dohi

Trends in Gastroenterology and Hepatology : Millennium 2000, Essential roles for Th2-type reponses in chronic intestinal inflammaton

胃肠病学和肝病学趋势：2000 年千年，Th2 型反应在慢性肠道炎症中的重要作用

• 期刊: Springer-Verlag
• 作者: Shirasawa S;T;Shirai Y;Kawamura T;Dohi T;Hirotomo Kato;Yuki I.Kawamura;Taeko Dohi;Kato H;Kawamura YI;Dohi T;Hashimoto M;Taeko Dohi;Kohtaro Fujihashi;Masahito Hashimoto;Masahito Hashimoto;Dohi T;Fujihashi K;Hashimoto M;Dohi T
• 通讯作者: Dohi T

Hirotomo Kato: "Lack of oral tolerance in aging is due to sequential loss of T cell responses in Peyer's patches"International Immunology. 15. 145-158 (2003)

Hirotomo Kato：“衰老过程中缺乏口服耐受性是由于派尔氏淋巴集结中 T 细胞反应连续丧失所致”《国际免疫学》。

Kawasaki, S.: "Intervention of thymus and activation-regulated chemokine attenuates the development of allergic airway inflammation and hvperresvonsiveness in mice"J Immunol. 166. 2055-2062 (2001)

Kawasaki, S.：“胸腺和活化调节趋化因子的干预可减轻小鼠过敏性气道炎症和高反应性的发展”JImmunol。