Expression of extra trinucleotide in CD44 variant of rheumatoid arthritis patients allows generation of disease-specific monoclonal antibody
Expression of extra trinucleotide in CD44 variant of rheumatoid arthritis patients allows generation of disease-specific monoclonal antibody
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DOI：
10.1016/j.jaut.2007.02.007
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发表时间：
2007-03-01
影响因子：
12.8
通讯作者：
Naor, David
中科院分区：
文献类型：
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作者：
Golan, Itshak;Nedvetzki, Shlomo;Naor, David
选择性靶向参与病理活动的细胞是医学研究的主要挑战。我们生成了单克隆抗体 (mAb)，其浓度范围为 2 至 100 μg/ml，专门结合类风湿性关节炎 (RA) 患者关节滑液细胞上表达的修饰 CD44 变体（称为 CD44vRA）。这些单克隆抗体仅在相对较高的浓度（200μg/ml）下与表达野生型CD44vRA（CD44v3-v10）的角质形成细胞发生交叉反应。 CD44vRA cDNA 的序列分析显示，43 名 RA 和银屑病关节炎患者中的 33 名患者体内存在额外的内含子衍生三核苷酸 CAG，它允许翻译额外的丙氨酸。这种插入使 RA 滑液细胞的细胞表面 CD44 发生构型变化，产生免疫原性表位并增强产生疾病特异性抗体的能力。事实上，抗 CD44vRA mAb（指定为 F8:33）能够诱导 RA 患者滑液细胞凋亡，但不能诱导同一患者的外周血白细胞、正常供体的角质形成细胞或骨关节炎患者的滑液细胞凋亡。此外，注射抗 CD44vRA mAb 可减少患有胶原诱导关节炎的 DBA/1 小鼠的关节炎症。这些发现表明抗 CD44vRA mAb 既具有生物活性又具有 RA 特异性。 (C) 2007 Elsevier Ltd. 保留所有权利。
Selective targeting of cells engaged in pathological activities is a major challenge for medical research. We generated monoclonal antibodies (mAbs) that exclusively bind, at concentrations ranging from 2 to 100 mu g/ml, to a modified CD44 variant (designated CD44vRA) expressed on synovial fluid cells from joints of rheumatoid arthritis (RA) patients. These mAbs cross-reacted with keratinocytes expressing wild type CD44vRA (CD44v3-v10) only at a relatively high concentration (200 mu g/ml). Sequence analysis of CD44vRA cDNA revealed, in 33 out of 43 RA and psoriatic arthritis patients, an extra intron-derived trinucleotide, CAG, which allows translation of an extra alanine. This insertion imposes a configurational change on the cell surface CD44 of RA synovial fluid cells, creating an immunogenic epitope and potentiating the ability to produce disease-specific antibodies. Indeed, the anti-CD44vRA mAbs (designated F8:33) were able to induce apoptosis in synovial fluid cells from RA patients, but not in peripheral blood leukocytes from the same patients, in keratinocytes from normal donors or in synovial fluid cells from osteoarthritis patients. Furthermore, injection of anti-CD44vRA mAbs reduced joint inflammation in DBA/1 mice with collagen-induced arthritis. These findings show that anti-CD44vRA mAbs are both bioactive and RA-specific. (C) 2007 Elsevier Ltd. All rights reserved.