van Lier R A, Borst J, Vroom T M, Klein H, Van Mourik P, Zeijlemaker W P, Melief C J
J Immunol. 1987 Sep 1;139(5):1589-96.
Two monoclonal antibodies (CLB-CD 27/1 and CLB-CD 27/2) were raised against a novel determinant on human T lymphocytes. One of these antibodies, CLB-CD 27/1 (clone 9F4), was grouped by the Third International Workshop and Conference on Human Leucocyte Differentiation Antigens together with three other monoclonal antibodies (VIT 14, OKT 18A, and S152) in the new cluster CD27. In this paper we show that antibodies belonging to this cluster recognize an antigen present on a large subset of peripheral T lymphocytes and most medullary thymocytes. At least two different nonoverlapping epitopes were identified with directly labeled monoclonal antibodies. Immunoprecipitation studies indicate that the target antigen of CD27 antibodies is a polypeptide of 55 kDa, which appears in the form of a disulfide-linked homodimer on the T lymphocyte membrane (Tp55). Stimulation of T cells via the T3/T cell antigen-receptor complex, with either phytohemagglutinin or CD3 monoclonal antibodies, resulted in a fivefold increase in the membrane expression of Tp55, whereas activation by phorbol myristate acetate caused a marked down-regulation. Moreover, an additional molecule of 32 kDa was precipitated from the membrane of activated but not of resting T cells. Addition of CD27 antibodies to cultures stimulated with either phytohemagglutinin or CD3 monoclonal antibody led to enhanced proliferation, whereas no effect was observed in phorbol myristate acetate or interleukin 2-stimulated cultures. The possible role of the Tp55 antigen in T cell activation is discussed.
制备了两种针对人T淋巴细胞上一种新决定簇的单克隆抗体(CLB-CD 27/1和CLB-CD 27/2)。其中一种抗体CLB-CD 27/1(克隆9F4),在第三届人类白细胞分化抗原国际研讨会和会议上,与其他三种单克隆抗体(VIT 14、OKT 18A和S152)一起被归入新的CD27簇。在本文中,我们表明属于该簇的抗体识别在外周T淋巴细胞的大部分和大多数髓质胸腺细胞上存在的一种抗原。用直接标记的单克隆抗体鉴定出至少两种不同的不重叠表位。免疫沉淀研究表明,CD27抗体的靶抗原是一种55 kDa的多肽,它以二硫键连接的同型二聚体形式出现在T淋巴细胞膜上(Tp55)。用植物血凝素或CD3单克隆抗体通过T3/T细胞抗原受体复合物刺激T细胞,导致Tp55的膜表达增加五倍,而佛波酯肉豆蔻酸酯激活则导致明显下调。此外,从活化但非静止的T细胞膜上沉淀出一种额外的32 kDa分子。将CD27抗体添加到用植物血凝素或CD3单克隆抗体刺激的培养物中导致增殖增强,而在佛波酯肉豆蔻酸酯或白细胞介素2刺激的培养物中未观察到作用。讨论了Tp55抗原在T细胞活化中的可能作用。