A monoclonal antibody recognizing human cancers with amplification/overexpression of the human epidermal growth factor receptor
A monoclonal antibody recognizing human cancers with amplification/overexpression of the human epidermal growth factor receptor
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DOI：
10.1073/pnas.232686499
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发表时间：
2003-01-21
影响因子：
11.1
通讯作者：
Old, LJ
中科院分区：
文献类型：
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作者：
Jungbluth, AA;Stockert, E;Old, LJ
表皮生长因子受体（EGFR）作为癌症治疗的靶点引起了广泛关注。野生型 (wt)EGFR 在多种肿瘤类型中扩增/过表达，并且已发现编码基因的几种突变形式，其中 AEGFR（缺乏外部结构域外显子 2-7 的缺失突变）是最常见的，尤其与胶质母细胞瘤相关。我们生成了针对 NR6(DeltaEGFR)（转染 DeltaEGFR 的小鼠成纤维细胞系 NR6）的单克隆抗体 (mAb)。在混合血液吸附测定、荧光激活细胞分选、蛋白质印迹和免疫组织化学中对 NR6(DeltaEGFR) 具有选择性反应性的 mAb 806 进行了详细分析，并与 mAb 528（抗 wtEGFR）和 DH8.3（抗 DeltaEGFR）进行了比较。在异种移植肿瘤和分子预分型的胶质母细胞瘤中，反应性模式如下：528与扩增和非扩增的wtEGFR反应； DH8.3 与 DeltaEGFR 反应； 806与扩增/过表达的wtEGFR（有或没有AEGFR）反应。在正常组织中，528 但 DH83 或 806 与许多器官广泛反应，例如表达高 EGFR 水平的肝脏。在胶质母细胞瘤和非 CNS 肿瘤组中，806 与高比例的胶质母细胞瘤以及大量肺癌和头颈上皮癌有反应。 DH8.3 反应性仅限于 DeltaEGFR 阳性胶质母细胞瘤。因此，806代表了一类mAb，它识别具有EGFR扩增/过度表达的肿瘤，但不识别正常组织或具有正常EGFR水平的肿瘤。我们的研究还表明 DeltaEGFR 仅限于胶质母细胞瘤，而其他报道则称这种突变存在于脑外肿瘤中。
Epidermal growth factor receptor (EGFR) has attracted considerable attention as a target for cancer therapy. Wild-type (wt)EGFR is amplified/overexpressed in a number of tumor types, and several mutant forms of the coding gene have been found, with AEGFR, a deletion mutation lacking exons 2-7 of the external domain, being the most common and particularly associated with glioblastoma. We generated monoclonal antibodies (mAbs) against NR6(DeltaEGFR) (mouse fibroblast line NR6 transfected with DeltaEGFR). mAb 806 with selective reactivity for NR6(DeltaEGFR) in mixed hemadsorption assays, fluorescence-activated cell sorting, Western blot, and immunohistochemistry was analyzed in detail and compared with mAbs 528 (anti-wtEGFR) and DH8.3 (anti-DeltaEGFR). In xenograft tumors and molecularly pretyped glioblastomas, the reactivity pattern was as follows: 528 reactive with amplified and nonamplified wtEGFR; DH8.3 reactive with DeltaEGFR; and 806 reactive with amplified/overexpressed wtEGFR (with or without AEGFR). In normal tissues, 528 but not DH83 or 806 was widely reactive with many organs, e.g., liver expressing high EGFR levels. In glioblastoma and non-CNS tumor panels, 806 was reactive with a high proportion of glioblastomas and a substantial number of epithelial cancers of lung and of head and neck. DH8.3 reactivity was restricted to DeltaEGFR-positive glioblastoma. Thus, 806 represents a category of mAbs that recognizes tumors with EGFR amplification/overexpression but not normal tissues or tumors with normal EGFR levels. Our study also indicates that DeltaEGFR is restricted to glioblastoma, in contrast to other reports that this mutation is found in tumors outside the brain.