Leitenberg D, Novak T J, Farber D, Smith B R, Bottomly K
Howard Hughes Medical Institute, Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
J Exp Med. 1996 Jan 1;183(1):249-59. doi: 10.1084/jem.183.1.249.
The CD45 tyrosine phosphatase plays an important role in regulating T lymphocyte activation, but the function of the different isoforms of CD45 is not known. T cell transfectants have been prepared that express individual CD45 isoforms in cells with a well-defined T cell receptor (TCR) from the D10 T helper 2 clone. We find that cells bearing low molecular weight CD45 isoforms are far more efficient in responding to stimulation with peptide and antigen-presenting cells compared with cells bearing high molecular weight CD45 isoforms. One hypothesis for the preferential activation of cells that express low molecular weight CD45 isoforms is that they interact with other cell surface antigens important in TCR signaling, altering their phosphorylation status and affecting the character of the signal transduction pathway. In this report, using cells expressing single isoforms, we demonstrate that low molecular weight isoforms of CD45 preferentially associate with CD4 and the TCR complex compared with high molecular weight isoforms. The molecular basis for this interaction was further examined using a glycosyl phosphatidyl inositol (GPI)-linked form of CD45Null (lacking tyrosine phosphatase domains), which preferentially associated with CD4 compared with GPI-linked CD45ABC, and cytoplasmic tail mutants of CD4, which retained the ability to coassociate. Using this panel of transfectants, it is clear that the interaction between CD4 and CD45 does not require the cytoplasmic domains of CD45, but is dependent on the specific external domain of the various isoforms: low molecular weight species were more likely to associate with the CD4-TCR complex than the higher molecular weight isoforms, and their ability to coassociate correlated with the magnitude of the response to specific antigen.
CD45酪氨酸磷酸酶在调节T淋巴细胞活化中起重要作用,但CD45不同异构体的功能尚不清楚。已制备了T细胞转染体,其在具有来自D10 T辅助2克隆的明确T细胞受体(TCR)的细胞中表达单个CD45异构体。我们发现,与携带高分子量CD45异构体的细胞相比,携带低分子量CD45异构体的细胞在对肽和抗原呈递细胞的刺激反应中效率要高得多。关于表达低分子量CD45异构体的细胞优先活化的一种假说是,它们与TCR信号传导中重要的其他细胞表面抗原相互作用,改变其磷酸化状态并影响信号转导途径的特性。在本报告中,使用表达单一异构体的细胞,我们证明与高分子量异构体相比,CD45的低分子量异构体优先与CD4和TCR复合物结合。使用糖基磷脂酰肌醇(GPI)连接形式的CD45Null(缺乏酪氨酸磷酸酶结构域)进一步研究了这种相互作用的分子基础,与GPI连接的CD45ABC相比,它优先与CD4结合,以及CD4的细胞质尾部突变体,其保留了共结合的能力。使用这组转染体,很明显CD4和CD45之间的相互作用不需要CD45的细胞质结构域,而是取决于各种异构体的特定外部结构域:低分子量物种比高分子量异构体更可能与CD4-TCR复合物结合,并且它们的共结合能力与对特定抗原的反应强度相关。