一种针对叶酸受体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及体内疗效

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

作者信息

Nagai Taku, Tanaka Masashi, Tsuneyoshi Yasuhiro, Matsushita Kakushi, Sunahara Nobuhiko, Matsuda Takemasa, Yoshida Hiroki, Komiya Setsuro, Onda Masanori, Matsuyama Takami

机构信息

Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.

出版信息

Arthritis Rheum. 2006 Oct;54(10):3126-34. doi: 10.1002/art.22082.

DOI:10.1002/art.22082

PMID:17009233

Abstract

OBJECTIVE

To investigate the effects of the recombinant immunotoxin dsFv anti-FRbeta-PE38, which consists of the disulfide-stabilized Fv fragment (dsFv) of the anti-folate receptor beta (anti-FRbeta) antibody and the 38-kd portion of Pseudomonas exotoxin A (PE38), on the activation and proliferation of cells that function in inflammatory and degradative processes in rheumatoid arthritis (RA) synovial tissue.

METHODS

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. The effects of dsFv anti-FRbeta-PE38 on the activation and proliferation of cells in RA synovial tissue were investigated by immunohistochemistry; the numbers of cells expressing CD68, vascular cell adhesion molecule 1, angiopoietin 1, CD34, proliferating cell nuclear antigen, and interleukin-6 and the numbers of apoptotic cells were counted in RA synovial tissue engrafted into SCID mice treated or not treated with dsFv anti-FRbeta-PE38. The effects of dsFv anti-FRbeta-PE38 on the generation of osteoclasts from RA adherent synovial mononuclear cells in vitro was investigated by counting the number of resorption pits on dentin slices treated or not treated with dsFv anti-FRbeta-PE38.

RESULTS

Administration of dsFv anti-FRbeta-PE38 reduced the numbers of macrophages, activated fibroblast-like cells, endothelial cells, and proliferating cells and increased the numbers of apoptotic cells in RA synovial tissue engrafted into SCID mice. In vitro, the generation of osteoclasts from RA adherent synovial mononuclear cells was largely suppressed by treatment with dsFv anti-FRbeta-PE38.

CONCLUSION

Our findings show that dsFv anti-FRbeta-PE38 immunotoxin would be a promising tool for the treatment of RA synovitis, especially when administered intraarticularly.

摘要

目的

研究重组免疫毒素dsFv抗FRβ-PE38对类风湿关节炎（RA）滑膜组织中参与炎症和降解过程的细胞激活和增殖的影响。该重组免疫毒素由抗叶酸受体β（抗FRβ）抗体的二硫键稳定Fv片段（dsFv）和铜绿假单胞菌外毒素A（PE38）的38kd部分组成。

方法

在大肠杆菌中产生Ig VH-PE38融合蛋白和Ig VL蛋白，然后通过对这些蛋白框架区域中的半胱氨酸残基进行工程改造，以二硫键连接。采用免疫组织化学方法研究dsFv抗FRβ-PE38对RA滑膜组织中细胞激活和增殖的影响；对移植到经或未经dsFv抗FRβ-PE38处理的SCID小鼠体内的RA滑膜组织中表达CD68、血管细胞黏附分子1、血管生成素1、CD34、增殖细胞核抗原和白细胞介素-6的细胞数量以及凋亡细胞数量进行计数。通过对经或未经dsFv抗FRβ-PE38处理的牙本质切片上的吸收凹坑数量进行计数，研究dsFv抗FRβ-PE38对体外RA贴壁滑膜单核细胞产生破骨细胞的影响。