THERAPEUTIC POTENTIAL OF ANTI-IL-6 RECEPTOR MONOCLONAL ANTIBODY FOR MURINE MODEL OF CROHN'S DISEASE

抗IL-6受体单克隆抗体对克罗恩病小鼠模型的治疗潜力

• 批准号: 10670469
• 负责人: ITO Hiroaki
• 金额: $ 2.05万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670469/
• 关键词: Crohn's disease; IL-6; IL-6 receptor; monoclonal antibody; wasting disease; IBD; CD45RB; Th1; クロ-ン病; IL-6レセプタ-; モノクロ-ナル抗体

To examine therapeutic potential of blocking IL-6 signaling for Crohn's disease, we introduced anti-IL-6 receptor monoclonal antibody (anti-IL-6R mAb) to a murine model of colitis. Colitis was induced in CB17-scid mice transferred CD45RBィイD1highィエD1 CD4ィイD1+ィエD1 T cells from normal Balb/c mice. Anti-IL-6R mAb or control rat IgG was administered intraperitoneally soon after T cell transfer, followed by weekly injection. Body weight was monitored weekly. Colons were removed at 8 weeks and colitis was graded histologically as 0 (minimal) to 3 (severe). ICAM-1 and VCAM-1 expression was analyzed by immunofluolescence staining and FACS, apoptotic cells were determined by TUNEL method. Mice treated with anti-IL-6R mAb showed normal growth while controls lost weight. Average colitis score was 0.64 for mAb treated mice, and 1.80 for controls. CD4ィイD1+ィエD1 T cells in the colon were apparently reduced by mAb as compared to massive infiltration in the controls. Colonic ICAM-1 and VCAM-1 expression was markedly suppressed by the treatment. In mice with colitis, ICAM-1 was expressed mainly in lamina propria, which was more remarkable in the luminal portion. ICAM-1ィイD1+ィエD1 cells were mostly Mac-1ィイD1+ィエD1 but not CD4ィイD1+ィエD1 cells. VCAM-1 was most strongly expressed by submucosal vascular endothelial cells. TUNELィイD1+ィエD1 apoptotic cells were very sparse despite massive infiltration of CD4ィイD1+ィエD1 T cells in colitic mice, while increased apoptosis was seen in mAb treated mice with reduced number of CD4ィイD1+ィエD1 T cells. These results suggest the therapeutic potential of anti-IL-6R mAb for human Crohn's disease.

为了研究阻断IL-6信号转导对克罗恩病的治疗潜力，我们将抗IL-6受体单抗(抗IL-6R单抗)引入小鼠结肠炎模型。将正常小鼠的CD45RbィイD1HighィエD1CD4CD4ィイD1+ィエD1T细胞转移至CB17SCID小鼠，诱导结肠炎。T细胞转移后即刻腹腔注射抗IL-6R单抗或对照大鼠免疫球蛋白，每周注射一次。每周监测体重。8周后切除结肠，组织学分级为0级(轻度)至3级(重度)。免疫荧光染色和流式细胞仪检测ICAM-1和VCAM-1的表达，TUNEL法检测细胞凋亡率。接受抗IL-6R单抗治疗的小鼠生长正常，而对照组小鼠体重下降。单抗治疗组小鼠平均结肠炎评分为0.64，对照组为1.80。与对照组相比，单抗组大鼠结肠内CD4CD4D1T细胞ィイ+ィエD1T细胞明显减少。治疗组大鼠结肠ICAM-1和VCAM-1的表达明显受抑制。在结肠炎小鼠中，ICAM-1主要在固有层表达，在管腔部分更为明显。ICAM-1CD4d1+ィイd1细胞多为ィエ-1ィイd1+ィエd1细胞，而不是ィイd1+ィエd1细胞。VCAM-1在黏膜下血管内皮细胞表达最强。尽管结肠炎小鼠体内有大量的ィイ-ィエ-D1+ィエ-D1T细胞，但TUNEL-ィイ-D1+-ィエ-D1T细胞的凋亡细胞非常稀少，而在单抗治疗的小鼠中，CD4-ィイ-D1+ィエ-D1T细胞数量减少，细胞凋亡率增加。这些结果提示抗IL-6R单抗对人类克罗恩病有治疗潜力。

项目成果

Ito H, et al: "Essential role for IL-6 in the pathogenesis of murine wasting disease and colitis."Gastroenterology. (in press).

Ito H 等人：“IL-6 在小鼠消耗性疾病和结肠炎发病机制中的重要作用。”胃肠病学。

Ito H, et al: "Anti-IL-6 receptor monoclonal antibody prevents Th1 cell-mediated murine colitis."GUT. 45 (Suppl V). A263 (1999)

Ito H 等人：“抗 IL-6 受体单克隆抗体可预防 Th1 细胞介导的小鼠结肠炎。”GUT。

Yamamoto M: "IL-6 is required for the development of Th1 cell-mediated murine colltis"J. Immunol.. (in press). (2000)

Yamamoto M：“IL-6 是 Th1 细胞介导的小鼠结肠炎发生所必需的”J.

Yamamoto M: "Anti-IL-6 receptor mAb prevents wasting disease-Th1 cell-mediated colitis in mice"Journal of Parenteral and Enteral Nutrition. 23(5). S163 (1999)

Yamamoto M：“抗 IL-6 受体单克隆抗体可预防小鼠消耗性疾病 - Th1 细胞介导的结肠炎”肠外和肠内营养杂志。

Ito H: "Inflammatory Bowel Diseases."Lippincot, Raven in press.

Ito H：“炎症性肠病”。Lippincot，Raven 正在出版。