Kolowos W, Gaipl U S, Voll R E, Frank C, Haas J P, Beyer T D, Kalden J R, Herrmann M
Institute of Clinical Immunology, Department of Internal Medicine III, Friedrich-Alexander-University, Erlangen-Nuremberg, Germany.
Lupus. 2001;10(5):321-31. doi: 10.1191/096120301671176280.
T cell activation was analysed in peripheral CD4+ T cells from both systemic lupus erythematosus (SLE) patients with active and inactive disease as well as in normal healthy donors (NHD) to investigate the involvement of CD4+ T cells in the etiopathogenesis of SLE. CD4+ T cell receptor (TCR) beta-chain transcripts, containing the complementarity determining region 3 (CDR3), were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and analysed by high-resolution polyacrylamide gel electrophoresis. In addition the CDR3 of both clonally activated as well as heterogeneous Vbeta families from SLE patients were analysed at the molecular level. We observed a restricted CDR3 length polymorphism in peripheral CD4+ T cells from SLE patients compared with NHD, more pronounced in patients with high disease activity. Furthermore, in some Vbeta families single peaks in the histogram indicated nearly monoclonal T cell expansion. Sequencing of selected TCR beta-chains revealed a increased content of acidic amino acids in the CDR3 encoded by either proximal Jbeta elements or N nucleotides. We conclude that CD4+ T cells from peripheral blood of SLE patients display features of a secondary antigen driven immune response. The bias of the CDR3 towards acidic amino acids suggests the involvement of positively charged antigens.
对患有活动期和非活动期系统性红斑狼疮（SLE）的患者以及正常健康供体（NHD）外周血中的CD4⁺ T细胞进行T细胞活化分析，以研究CD4⁺ T细胞在SLE发病机制中的作用。通过逆转录聚合酶链反应（RT-PCR）扩增包含互补决定区3（CDR3）的CD4⁺ T细胞受体（TCR）β链转录本，并通过高分辨率聚丙烯酰胺凝胶电泳进行分析。此外，还在分子水平上分析了SLE患者克隆激活的和异质性Vβ家族的CDR3。我们观察到，与NHD相比，SLE患者外周血CD4⁺ T细胞中CDR3长度多态性受限，在疾病活动度高的患者中更为明显。此外，在一些Vβ家族中，直方图中的单峰表明T细胞几乎呈单克隆扩增。对选定的TCR β链进行测序发现，由近端Jβ元件或N核苷酸编码的CDR3中酸性氨基酸含量增加。我们得出结论，SLE患者外周血中的CD4⁺ T细胞表现出二次抗原驱动免疫反应的特征。CDR3向酸性氨基酸的偏向表明带正电荷的抗原参与其中。
