A preclinical study using recombinant anti-folate receptor-beta immunotoxin for the treatment of rheumatoid arthritis

重组抗叶酸受体-β免疫毒素治疗类风湿性关节炎的临床前研究

• 批准号: 17591051
• 负责人: MATSUYAMA Takami
• 金额: $ 2.37万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591051/
• 关键词: Immunotoxin; Folate receptor; Rheumatoid arthritis; SCID; Macrophages; Activated fibroblast-like cells; Osteoclasts; SCID

OBJECTIVE: To investigate the effects of the recombinant immunotoxin dsFv anti-folate receptor beta (FRβ) on the activation and proliferation of the cell that function in inflammatory and degradative processes in rheumatoid arthritis synovial tissue.20051) Productior of recombinant type anti-human FRβ immunotoxin: The Ig VH-PE38 fusion protein and the Ig VL protein were produced in Escherichia coli, and then joined with a disulfide bond by engineering cysteine residues in the framework regions of these proteins.2) In vitro study: The immunotoxin significantly induced the apoptosis of FRβ-transfected macrophages and adherent RA synovial mononuclear cells.3) In vivo study: Administration of immunotoxin reduced the numbers of CD68^+ macrophages, activated fibroblast-like cells (VCAM-1^+, Angiopoietin-1^+), inflammatory cytokines (TNF-α, IL-6), angiogenesis (CD34^+ cells) in RA synovial tissue engrafted into SCID mice.20061) Tissue localization of FRβ in rhesus monkey: The liver and skin weakly expressed FRβ in rhesus monkey.2) Productior of rat-anti mouse FRβ monoclonal antibody: Murine FRβ cDNA was transfected into rat mastcytorra, and injected into foot-pad of WKY rats. Nine monoclonal antibodies were obtained. Any monoclonal antibodies were not cross-reacted with mouse FRα.3) Tissue localization of FRβ in mouse: FRβ were expressed in thioglycolate-induced peritoneal macrophages (++++), liver (+ 〜 ++), lung (+ 〜 ++) and colon (+), suggesting that inflammatory stimulation may up-regulate; FRβ^+ cells in mouse and rhesus monkey.Conclusion: Immunotoxin would be a promising tool for the treatment of RA. Prospects: Now, investigators are producing anti-mouse FRβ immunotoxin and will examine the effects on murine arthritis model (CIIA-induced arthritis and serum transfer-induced arthritis).

目的：探讨重组免疫毒素dsFv抗叶酸受体β（FRβ）对类风湿性关节炎滑膜组织炎症和降解过程中细胞活化和增殖的影响。20051）重组型抗人FRβ免疫毒素的产生：Ig VH-PE38融合蛋白和Ig VL 蛋白质是在大肠杆菌中产生的，然后通过在这些蛋白质的框架区改造半胱氨酸残基来连接二硫键。2) 体外研究：免疫毒素显着诱导 FRβ 转染的巨噬细胞和贴壁 RA 滑膜单核细胞的凋亡。3) 体内研究：免疫毒素的施用减少了 移植到 SCID 小鼠的 RA 滑膜组织中的 CD68^+ 巨噬细胞、活化的成纤维细胞样细胞（VCAM-1^+、Angiopoietin-1^+）、炎症细胞因子（TNF-α、IL-6）、血管生成（CD34^+ 细胞）。20061) 恒河猴中 FRβ 的组织定位：肝脏和皮肤中 FRβ 弱表达 2)大鼠抗小鼠FRβ单克隆抗体的制备：将鼠FRβcDNA转染大鼠肥大细胞，注射至WKY大鼠足垫。获得了九种单克隆抗体。任何单克隆抗体均不与小鼠FRα发生交叉反应。3）FRβ在小鼠体内的组织定位：FRβ在巯基乙酸诱导的腹腔巨噬细胞（++++）、肝脏（+〜++）、肺（+〜++）和结肠（+）中表达，提示炎症刺激可能上调；小鼠和恒河猴中的FRβ^+细胞。结论：免疫毒素将是治疗RA的有前途的工具。前景：现在，研究人员正在生产抗小鼠FRβ免疫毒素，并将检查其对小鼠关节炎模型（CIIA诱导的关节炎和血清转移诱导的关节炎）的影响。

项目成果

Effectiveness of Anti-Folate Receptor β Antibody Conjugated With Truncated Pseudomonas Exotoxin in the Targeting of Rhematoid Arthritis Synovial Macrophages

抗叶酸受体 β 抗体与截短假单胞菌外毒素缀合在类风湿性关节炎滑膜巨噬细胞靶向中的有效性

• 发表时间: 2005
• 期刊: Arthritis and Rheumatism 52
• 作者: Li S-M.;Li G-M.;Nakamura S.;Ikuta K.;Nakaya T.;Nagayoshi R et al.
• 通讯作者: Nagayoshi R et al.

In vitro and in vivo efficacy of a recombinant immunotoxin against folate receptor beta on the activation and proliferation of rheumatoid arthritis synovial cells.

针对叶酸受体β的重组免疫毒素对类风湿性关节炎滑膜细胞活化和增殖的体外和体内功效。

• 发表时间: 2006
• 期刊: Arthritis ＆ Rheumatism 54(10)
• 作者: Nagai T;Tanaka M;Tsuneyoshi Y;Matsushita K;Sunahara N;Matsuda T;Yoshida H;Komiya S;Onda M;Matsuyama T.
• 通讯作者: Matsuyama T.