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由一种急性淋巴细胞白血病细胞系引发的抗MHC II类单克隆抗体的补充特性:免疫荧光细胞荧光测定法、补体依赖性细胞毒性、抗原的二维分析

Supplementary characteristics of anti-MHC class II monoclonal antibodies elicited by an ALL cell line: immunofluorescence cytofluorometry, C-dependent cytotoxicity, two-dimensional analysis of antigen.

作者信息

Chorváth B, Duraj J, Sedlák J, Plesková I, Munozová H, Buc M

机构信息

Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Czechoslovakia.

出版信息

Neoplasma. 1987;34(4):417-25.

PMID:3309687
Abstract

Monoclonal antibodies directed to MHC class II antigen(s), elicited by a non-T, non-B ALL cell line, were characterized by immunofluorescence flow cytofluorometry and ELISA immunofiltration measurements of their immunoreactivity with selected neoplastic hemopoietic cell lines, determination of their complement-dependent cytotoxic activity against isolated peripheral blood B and T lymphocytes and by two-dimensional electrophoretic analysis (isoelectric focusing, SDS-PAGE) of radiolabeled, immunoprecipitated by these antibodies cell surface antigens. Patterns of these immunological reactivities, as well as two-dimensional radioimmunoprecipitation patterns (acidic heavy chain p35 and basic light chain p30) of antigens recognized by these antibodies confirm their anti-MHC class II specificity. One of these antibodies (braFB6; IgG2b) displayed identical pattern of expression on cell lines and cell types as the typical anti-MHC class II antibodies, but immunoprecipitated only a single chain p30 radioiodinated cell surface protein (with two-dimensional pattern close to the beta-chain of MHC class II DR antigen). These properties indicate the ability of braFB6 monoclonal antibody to recognize a nonpolymorphic determinant of DP-MHC class II antigen.

摘要

由一种非T、非B急性淋巴细胞白血病细胞系引发的、针对MHCⅡ类抗原的单克隆抗体,通过免疫荧光流式细胞荧光测定法以及ELISA免疫过滤法对其与选定的肿瘤造血细胞系的免疫反应性进行了表征,测定了其对分离的外周血B淋巴细胞和T淋巴细胞的补体依赖性细胞毒活性,并通过二维电泳分析(等电聚焦、SDS-PAGE)对经这些抗体免疫沉淀的放射性标记细胞表面抗原进行了分析。这些免疫反应模式以及这些抗体识别的抗原的二维放射免疫沉淀模式(酸性重链p35和碱性轻链p30)证实了它们的抗MHCⅡ类特异性。其中一种抗体(braFB6;IgG2b)在细胞系和细胞类型上表现出与典型抗MHCⅡ类抗体相同的表达模式,但仅免疫沉淀了一种单链p30放射性碘化细胞表面蛋白(二维模式接近MHCⅡ类DR抗原的β链)。这些特性表明braFB6单克隆抗体能够识别DP-MHCⅡ类抗原的一个非多态性决定簇。

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