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用葡萄球菌蛋白A分析细胞表面标志物:对哺乳动物淋巴决定簇的荧光研究

Analysis of cell-surface markers with staphylococcal protein A: fluorescence studies on mammalian lymphoid determinants.

作者信息

Dorval G, Wigzell H, Leibold W, Killander D

出版信息

J Immunol Methods. 1976;9(3-4):251-66. doi: 10.1016/0022-1759(76)90200-3.

Abstract

Membrane antigens including different classes of immunoglobulins, transplantation antigens, beta2-microglobulin, T lymphocyte specific antigens, and virally determined surface components were investigated using fluorescein-labeled Staphylococcal protein A in combination with cytofluorometric studies. Lymphocytes of seven species: mouse, rat, guinea pig, pig, cow, monkey, and human, and of ten human lymphoma-derived lines were tested. Analysis of the differential expression of surface markers revealed a reproducible reaction of protein A with cell-surface Fc of IgG actively produced by lymphoid cells from human, monkey, guinea pig, and pig, and with passively attached IgG molecules in the form of antibodies, directed against cell surface antigens of all lymphoid cells tested. No surface Ig was detected on so-called T lymphocytes. The distribution of cell-bound Ig density among surface Ig-positive cells was found to be different depending upon the origin of the cells with regard to lymphoid organ; it was parallel among the lymphoma lines tested and on peripheral blood cells from human, monkey, and pig, although large variations in fluorescence intensity among individual cells and among the different lines were recorded. Beta2-microglobulin determinants were found equally well on enriched human T and B cells. Transplantation, and T lymphocyte-specific antigens were detected on the majority of the lymphoid cells and on a restricted population respectively.

摘要

利用荧光素标记的葡萄球菌蛋白A结合细胞荧光测定研究,对包括不同类别的免疫球蛋白、移植抗原、β2微球蛋白、T淋巴细胞特异性抗原以及病毒决定的表面成分在内的膜抗原进行了研究。测试了七种物种(小鼠、大鼠、豚鼠、猪、牛、猴和人)的淋巴细胞以及十种人淋巴瘤衍生细胞系。对表面标志物差异表达的分析显示,蛋白A与人、猴、豚鼠和猪的淋巴细胞主动产生的细胞表面IgG的Fc以及以抗体形式被动附着的IgG分子(针对所有测试淋巴细胞的细胞表面抗原)发生可重复反应。在所谓的T淋巴细胞上未检测到表面Ig。发现表面Ig阳性细胞中细胞结合Ig密度的分布因淋巴细胞来源的淋巴器官不同而有所差异;在所测试的淋巴瘤细胞系以及人、猴和猪的外周血细胞中,其分布是平行的,尽管记录到单个细胞之间以及不同细胞系之间荧光强度存在很大差异。在富集的人T细胞和B细胞上同样能很好地检测到β2微球蛋白决定簇。移植抗原和T淋巴细胞特异性抗原分别在大多数淋巴细胞和有限的细胞群体上被检测到。

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