Treatment for rheumatoid arthritis with immunogene therapy by angiostatin

血管抑制素免疫基因疗法治疗类风湿性关节炎

• 批准号: 13670486
• 负责人: SAITO Kazuyoshi
• 金额: $ 2.37万
• 依托单位: University of Occupational and Environmental Health
• 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 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670486/
• 关键词: rheumatoid arthritis; gene therapy; monoclonal antibody; angiostatin

The aim of this research is establishment of antigen-specific immunogene therapy for rheumatoid arthritis with angiostatin. At first, we combined IL-10 expression vector and monoclonal antibodies against human ICAM-1 using sulfo-succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo-SMCC). Then we added the conjugates to ICAM-1 expressing cells (ICAM-1 COS ; COS cells stably trasfected with ICAM-1 expression vector, synovial fibroblast cell line E11). We detected significant IL-10 in the supernatant of ICAM-1 expressing cells but not in control COS cells after 72hour incubation with immunogene. The tranfection efficiency increased four times when the expression vector was conjugated to Fab fragment of antibody. The addition of native antibody to the medium reduced transfection efficiency, supporting antigen-specific gene delivery of this method. We are planning to conjugate angiostatin gene and ICAM-1 monoclonal antibody and infuse to RA SCID mice (SCID mice transplanted with human synovial tissue)

本研究的目的是建立血管抑素对类风湿关节炎的抗原特异性免疫基因治疗。首先，我们利用磺基琥珀酰亚胺基4-（N-马来酰亚胺甲基）环己烷-1-羧酸酯（sulfo-succinimidyl 4-（N-maleimidomethyl）hexane-1-carboxylic acid，sulfo-SMCC）将IL-10表达载体与抗人ICAM-1单克隆抗体结合。然后，我们将缀合物加入表达ICAM-1的细胞（ICAM-1 COS ;用ICAM-1表达载体稳定转染的COS细胞，滑膜成纤维细胞系E11）。免疫组化法检测到ICAM-1表达细胞培养上清中IL-10的表达，而对照COS细胞培养上清中IL-10的表达无明显变化。当表达载体与抗体Fab段偶联时，转染效率提高了4倍。向培养基中添加天然抗体降低了转染效率，支持该方法的抗原特异性基因递送。我们计划将血管抑素基因与ICAM-1单克隆抗体偶联，并注入RA SCID小鼠（移植人滑膜组织的SCID小鼠）

项目成果

Nakayamada, S. et al.: "β1 integrin-mediated signaling induces intercellular adhesion molecule 1 and Fas on rheumatoid synovial cells and Fas-mediated apoptosis"Arthritis and Rheum.. 48:(5). 1239-1248 (2003)

Nakayamada，S.等人：“β1整联蛋白介导的信号传导诱导类风湿滑膜细胞上的细胞间粘附分子1和Fas以及Fas介导的细胞凋亡”Arthritis and Rheum.. 48：(5)(2003)。

Nakayamada, S.: "β1 integrin-mediated signaling induces intercellular adhesion molecule 1 and Fas on rheumatoid synovial cells and Fas-mediated apoptosis"Arthritis and Rheum.. 48:(5). 1239-1248 (2003)

Nakayamada, S.：“β1 整合素介导的信号传导诱导类风湿滑膜细胞上的细胞间粘附分子 1 和 Fas 以及 Fas 介导的细胞凋亡”Arthritis and Rheum.. 48：(5) (2003)。

Nakayamada, S.: "β1 Integrin/Focal Adhesion Kinase-mediated Signaling Induces Intercellular Adhesion Moledule 1 and Receptor Activator of Nuclear Factor κB Ligand on Osteoblasts and Osteoclast Maturation"JBC. 278:(46). 45368-45374 (2003)

Nakayamada, S.：“β1 整合素/粘着斑激酶介导的信号传导诱导成骨细胞和破骨细胞成熟的细胞间粘着分子 1 和核因子 κB 配体的受体激活剂”JBC 278：(46)。

Saito, K.: "Succeessfurl treatment with anti-CD20 monoclonal antibody (rituximab) of life-threatening refratory systemic lupus erythematosus with renal and central nervous system involvement"LUPUS. 12. 798-800 (2003)

Saito, K.：“用抗 CD20 单克隆抗体（利妥昔单抗）成功治疗累及肾脏和中枢神经系统的危及生命的难治性系统性红斑狼疮”狼疮。

Nakayamada, S. et al.: "β Integrin/Focal Adhesion Kinase-mediated Signaling Induces Intercellular Adhesion Molecule 1 and Receptor Activator of Nuclear Factor κB Ligand on Osteoblasts and Osteoclast Maturation"JBC. 278(46). 45368-45374 (2003)

Nakayamada，S.等：“β整合素/粘着斑激酶介导的信号传导诱导成骨细胞和破骨细胞成熟的细胞间粘着分子1和核因子κB配体的受体激活剂”JBC 278(46)。