单克隆抗角蛋白抗体：制备、特性及免疫组织化学应用

Monoclonal antikeratin antibody: production, characterization, and immunohistochemical application.

作者信息

Eto H, Hashimoto K, Kobayashi H, Matsumoto M, Kanzaki T, Mehregan A H, Weiss R A

出版信息

J Invest Dermatol. 1985 May;84(5):404-9. doi: 10.1111/1523-1747.ep12265506.

PMID:2582061

Abstract

A monoclonal antikeratin antibody, EKH4, was produced from a hybridoma cell line which was established by fusing P3X63SAg8 mouse myeloma cells with spleen cells of mice immunized with human trichilemmoma cells. Immunoblot analysis showed that EKH4 antibody reacts predominantly with 50 kilodalton keratin polypeptide in normal epidermis. By indirect immunofluorescence and immunoperoxidase techniques, EKH4 antibody reacted with the lower 2-3 cell layers of the epidermis as well as most cells of pilosebaceous follicle of human and animal skin. Tumor cells of human basal cell epitheliomas and squamous cell carcinomas were also stained with this antibody. The staining was much more regular and intense compared with an available monoclonal antikeratin antibody, AE1. In the lesion of epidermal proliferative disorders, such as psoriasis and actinic keratosis, the entire epidermis instead of the lower layers was stained with EKH4 antibody. Normal skin overlying or adjacent to epithelial tumors also showed positive staining in the entire epidermis. By using indirect immunoperoxidase technique, EKH4 also stained alcohol-fixed, paraffin-embedded tissue sections.

摘要

一种单克隆抗角蛋白抗体EKH4是由一个杂交瘤细胞系产生的，该细胞系是通过将P3X63SAg8小鼠骨髓瘤细胞与用人毛鞘瘤细胞免疫的小鼠脾细胞融合而建立的。免疫印迹分析表明，EKH4抗体在正常表皮中主要与50千道尔顿的角蛋白多肽发生反应。通过间接免疫荧光和免疫过氧化物酶技术，EKH4抗体与表皮的下2 - 3层细胞以及人和动物皮肤的毛囊的大多数细胞发生反应。人基底细胞上皮瘤和鳞状细胞癌的肿瘤细胞也被该抗体染色。与现有的单克隆抗角蛋白抗体AE1相比，这种染色更加规则和强烈。在表皮增生性疾病如银屑病和光化性角化病的病变中，EKH4抗体染色的是整个表皮而非下层。上皮肿瘤上方或相邻的正常皮肤在整个表皮中也显示出阳性染色。通过使用间接免疫过氧化物酶技术，EKH4也能对酒精固定、石蜡包埋的组织切片进行染色。