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Significance of anti-calpastatin antibodies in rheumatic diseases and their effect on osteoclast

抗钙蛋白酶抑制剂抗体在风湿性疾病中的意义及其对破骨细胞的影响

基本信息

批准号：
12670437
负责人：
MIMORI Tsuneyo
金额：
$ 2.05万
依托单位：
KYOTO UNIVERSITY (2001-2002)Keio University (2000)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

I. Role of calpain in osteoclast activity and effect of anti-calpastatin antibodies in rheumatic diseases:Osteoclasts prepared from newborn rabbit femur were caltured on the thin-sliced ivory, and the effect of calpain on the bone absorption of osteoclasts was examined. The bone absorption activity decreased up to 50% of control by adding human calpastatin and calpain inhibitory peptide and increased 37-45% by mouse monoclonal anti-calpastatin antibodies. IgG from some RA patient sera with anti-calpastatin antibodies also increased the activity up to 32%. Thus, calpain/calpastatin system affects the osteoclast activity and may be associated with joint destruction in RA patients.II. Effect of calpain inhibitors on anti-collagen monoclonal antibody-induced mouse arthritis:Since calpain is thought to be a neutral proteinase that affects cartilage destruction and inflammation, the inhibition of calpain activity might be a new therapeutic strategy of RA. Therefore, we intended to treat RA model mouse by calpain inhibitors. Various calpain inhibitors (calpastatin, leupeptin, calpeptin and E-64-d) inhibited the development of anti-collagen monoclonal antibody cocktail-induced mouse arthritis, and in particular the high dose E-64-d (9mg/kg) showed the significant inhibition of arthritis as strong as dexamethasone.III. Development of quatitative mesurement of anti-calpastatin antibodies in rheumatic dseases:We previously reported that autoantibodies to calpastatin were detected in patients with rhuematic diseases. In this study, quantitative ELISA was developed by using purified human recombinant calpastatin that was expressed from the full-length cDNA encoding for human calpastatin. Anti-calpastatin antibodies were detected in RA with the highest frequency (43%), whereas also detected in SLE (30%), scleroderma (30%) and PM/DM (24%).

一、钙蛋白酶在破骨细胞活性中的作用及抗钙蛋白酶抗体在风湿性疾病中的作用：取新生兔股骨制备破骨细胞，在薄片象牙上培养，观察其对破骨细胞骨吸收的影响。加入人Calastatin和Calain抑制肽后，骨吸收活性降低达对照组的50%，而加入鼠抗Calastatin单抗后，骨吸收活性增加37~45%。一些类风湿关节炎患者血清中的抗钙调蛋白抗体的免疫球蛋白也使活性提高了32%。因此，calain/calastatin系统影响破骨细胞的活性，并可能与RA患者的关节破坏有关。ii.calain抑制剂对抗胶原单抗诱导的小鼠关节炎的影响：由于calain被认为是一种影响软骨破坏和炎症的中性蛋白酶，因此抑制calain活性可能成为RA的一种新的治疗策略。因此，我们打算用钙蛋白酶抑制剂治疗类风湿关节炎模型小鼠。不同的钙蛋白酶抑制剂(钙粘蛋白、亮肽素、钙肽蛋白和E--d)可抑制抗胶原单抗鸡尾酒诱导的小鼠关节炎的发展，尤其是大剂量的E--d(9 mg/kg)对关节炎的抑制作用与地塞米松一样强。III.风湿性疾病中抗钙蛋白酶抗体定量检测的进展：我们先前报道在风湿性疾病患者中检测到抗钙蛋白酶的自身抗体。在本研究中，利用从编码人钙蛋白酶抑素的全长cDNA中表达的纯化的重组人钙蛋白酶抑素建立了定量酶联免疫吸附试验。抗Calastatin抗体在RA中的阳性率最高(43%)，在SLE(30%)、硬皮病(30%)和PM/DM(24%)中也有阳性。