Expression of the sialosyl-Tn epitope on CD45 derived from activated peripheral blood T cells
Expression of the sialosyl-Tn epitope on CD45 derived from activated peripheral blood T cells
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DOI：
10.3109/08820139809022707
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发表时间：
1998-01-01
影响因子：
2.8
通讯作者：
Winfield, J
中科院分区：
文献类型：
--
作者：
Fernsten, P;Shaw, M;Winfield, J
细胞表面蛋白酪氨酸磷酸酶CD 45是系统性红斑狼疮（SLE）中IgM抗T细胞自身抗体的主要靶点。CD 45上的自身反应性决定簇是在活化的T细胞和某些T细胞系上表达的O-连接聚糖，而不是线性或构象多肽表位或N-连接聚糖。为了鉴定可能在CD 45的功能性相互作用中起作用或者是SLE抗CD 45自身抗体的候选靶表位的寡糖结构，通过ELISA测试从Jurkat T细胞纯化的自身反应性CD 45和从CLL B细胞纯化的非自身反应性CD 45的粘蛋白型O-聚糖结构的表达。针对血型A、1型H链、2型H链、T、Le（a）、唾液酸化-Lee、Le B、唾液酸化-Le（c）、Lex、唾液酸化-Le（x）、多岩藻糖基化Le（x）、Le（y）和唾液酸化-延伸Le（y）的单克隆抗体（mAb）均不能与来自B细胞或T细胞的CD 45反应。然而，针对Tn（galNAc α 1-O-ser/thr）或唾液酸-Tn（neuNAc α 2-GgalNAc α 1-O-ser/thr）结构的mAb与源自Jurkat T细胞的CD 45反应，但不与源自CLL B细胞的CD 45反应。在蛋白质印迹法中，抗Tn mAb还与从Jurkat T细胞分离的CD 45反应，但不与从GEM、MOLT-3或PEER T细胞; Daudi、Raji或CLL B细胞;或静息或Con A激活的PBL分离的CD 45反应。然而，抗唾液酸-Tn单克隆抗体染色从Jurkat和CEM T细胞和Con A激活的PBL分离的CD 45的印迹，反应性模式类似于抗CD 45自身抗体。流式细胞术分析表明，唾液酸-Tn表位表达的CD 4 +/CD 8- T细胞的亚群。
The cell surface protein tyrosine phosphatase CD45 is a major target of IgM anti-T cell autoantibodies in systemic lupus erythematosus (SLE). The autoreactive determinants on CD45 are O-linked glycans expressed on activated T cells and certain T cell lines, rather than linear or conformational polypeptide epitopes or N-linked glycans. To identify oligosaccharide structures that may play a role in the functional interactions of CD45 or are candidate target epitopes of SLE anti-CD45 autoantibodies, autoreactive CD45 purified from Jurkat T cells and non-autoreactive CD45 purified from CLL B cells were tested by ELISA for expression of mucin-type O-glycan structures. Monoclonal antibodies (mAbs) directed against blood group A, type 1 H chains, type 2 H chains, T, Le(a), sialylated-lee, Leb, sialylawted-Le(c), Lex, sialylated-Le(x), multi-fucosylated Le(x), Le(y), and sialylated-extended Le(y) failed to react with CD45 from either B cells or T cells. However, mAbs directed against Tn (galNAc alpha 1-O-ser/thr) or sialosyl-Tn (neuNAc alpha 2-GgalNAc alpha 1-O-ser/thr) structures reacted with CD45 derived from Jurkat T cells, but not from CLL B cells. Anti-Tn mAbs also reacted in western blotting procedures with CD45 isolated from Jurkat T cells, but did not react with CD45 isolated from GEM, MOLT-3, or PEER T cells; Daudi, Raji, or CLL B cells; or resting or Con A-activated PBL. However, anti-sialosyl-Tn mAbs stained blots of CD45 isolated from Jurkat and CEM T cells and Con A-activated PBL, a pattern of reactivity similar to that of the anti-CD45 autoantibodies. Flow cytometric analyses demonstrated that the sialosyl-Tn epitopes are expressed on a subpopulation of CD4+/CD8- T cells.