Stross W P, Warnke R A, Flavell D J, Flavell S U, Simmons D, Gatter K C, Mason D Y
Nuffield Department of Pathology, University of Oxford, John Radcliffe Hospital.
J Clin Pathol. 1989 Sep;42(9):953-61. doi: 10.1136/jcp.42.9.953.
Three monoclonal antibodies MT1, L60 (Leu-22), and DF-T1, were reported independently as recognising human T cells in routinely processed, paraffin wax embedded tissue. The present study was performed to compare these three reagents in terms of their immunocytochemical reactions and target molecule(s). On Western blotting of white cell extracts the three antibodies reacted with antigens of the same molecular weight (range 110-160 kilodaltons). Furthermore, their immunocytochemical reactivity with normal human cells, as analysed by two-colour flow cytometry, was essentially identical (labelling of monocytes, most T lymphocytes, and weak reactions with some B cells), and the antibodies gave closely similar reactions on 54 white cell derived neoplasms. To identify the target antigen for these three reagents, antibodies from the Third International Workshop on Leucocyte Antigens were reviewed and it was shown that the Western blotting and immunocytochemical reactions of MT1, L60 (Leu-22), and DF-T1 were identical with those of the reagents which defined the CD43 antigen (also known as leucosialin or sialophorin). Furthermore, all these antibodies reacted with cells transfected with a cDNA clone encoding CD43. It is concluded that antibodies MT1, L60 (Leu-22), and DF-T1 all recognise the heavily glycosylated myeloid/lymphoid associated CD43 antigen.
三种单克隆抗体MT1、L60(Leu-22)和DF-T1曾被分别报道可识别常规处理的石蜡包埋组织中的人T细胞。本研究旨在比较这三种试剂在免疫细胞化学反应和靶分子方面的情况。在白细胞提取物的蛋白质印迹分析中,这三种抗体与分子量相同(范围为110 - 160千道尔顿)的抗原发生反应。此外,通过双色流式细胞术分析,它们与正常人细胞的免疫细胞化学反应基本相同(标记单核细胞、大多数T淋巴细胞,与一些B细胞有弱反应),并且这三种抗体对54种白细胞来源的肿瘤细胞产生非常相似的反应。为了确定这三种试剂的靶抗原,查阅了第三届白细胞抗原国际研讨会的抗体,结果显示MT1、L60(Leu-22)和DF-T1的蛋白质印迹和免疫细胞化学反应与定义CD43抗原(也称为白细胞唾液酸蛋白或涎福林)的试剂相同。此外,所有这些抗体都与转染了编码CD43的cDNA克隆的细胞发生反应。得出的结论是,抗体MT1、L60(Leu-22)和DF-T1均识别高度糖基化的髓系/淋巴系相关CD43抗原。