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单克隆抗体45-2D9可识别一种人c-Ha-ras转化细胞系(45-342)上的细胞表面糖蛋白以及人类肿瘤上的一个共同表位。

Monoclonal antibody 45-2D9 recognizes a cell surface glycoprotein on a human c-Ha-ras transformed cell line (45-342) and a shared epitope on human tumors.

作者信息

Roth J A, Ames R S, Restrepo C, Scuderi P

出版信息

J Immunol. 1986 Oct 1;137(7):2385-9.

PMID:3531330
Abstract

A monoclonal antibody (45-2D9) produced after immunization of BALB/c mice with the c-Ha-ras NIH 3T3 tertiary transfectant (45-342) recognized a determinant expressed by the primary, three of three secondary, and one of three tertiary transfectants, but not by NIH 3T3 cells. The determinant was present on the cell surface and was distinct from murine leukemia virus gp70 by absorption studies. Biosynthetic labeling and immunoprecipitation studies with [35S]methionine and [3H]glucosamine demonstrated that 45-2D9 recognizes a 74,000 Mr glycoprotein with minor bands of 90,000 and 180,000 Mr on SDS-PAGE. Pulse chase studies demonstrated a 68,000 Mr precursor molecule that incorporated only [35S]methionine. The distribution of the epitope recognized by 45-2D9 was assessed by immunoperoxidase staining. The antigen was not detected on 10 primary and metastatic murine tumors or 11 transformed murine cell lines. However, a variety of surgically excised human tumors demonstrated intense staining, whereas staining of normal tissues was minimal or not detectable. Thus a human oncogene-transfected cell can express a new cell surface determinant apparently unrelated to the oncogene product, which is also selectively expressed by human tumors.

摘要

用c-Ha-ras NIH 3T3三级转染细胞(45-342)免疫BALB/c小鼠后产生的单克隆抗体(45-2D9)识别一种由原代转染细胞、三个二代转染细胞中的三个以及三个三代转染细胞中的一个所表达的决定簇,但不被NIH 3T3细胞识别。通过吸收研究表明,该决定簇存在于细胞表面,且与鼠白血病病毒gp70不同。用[35S]甲硫氨酸和[3H]葡糖胺进行的生物合成标记和免疫沉淀研究表明,45-2D9在SDS-PAGE上识别一种74,000 Mr的糖蛋白,还有90,000和180,000 Mr的次要条带。脉冲追踪研究表明存在一个仅掺入[35S]甲硫氨酸的68,000 Mr前体分子。通过免疫过氧化物酶染色评估45-2D9识别的表位分布。在10个原发性和转移性鼠肿瘤或11个转化的鼠细胞系上未检测到该抗原。然而,各种手术切除的人类肿瘤显示出强烈染色,而正常组织的染色则很轻微或无法检测到。因此,一个人癌基因转染细胞可以表达一种显然与癌基因产物无关的新细胞表面决定簇,该决定簇也在人类肿瘤中选择性表达。

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