Reconstitution of deficient T cell receptor ζ chain restores T cell signaling and augments T cell Receptor/CD3-induced interleukin-2 production in patients with systemic lupus erythematosus
Reconstitution of deficient T cell receptor ζ chain restores T cell signaling and augments T cell Receptor/CD3-induced interleukin-2 production in patients with systemic lupus erythematosus
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DOI：
10.1002/art.11072
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发表时间：
2003-07-01
影响因子：
--
通讯作者：
Tsokos, GC
中科院分区：
文献类型：
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作者：
Nambiar, MP;Fisher, CU;Tsokos, GC
目标。大多数系统性红斑狼疮(SLE)患者的T细胞表现出与T细胞受体(TCR)X1链缺陷相关的抗原受体介导的信号异常，X1链是TCR/CD3复合体的亚单位。本研究旨在探讨强迫表达TCR链是否可能逆转SLE T细胞已知的信号异常和白介素2(IL-2)产生缺陷。利用最近发展起来的核孔化技术，将构建在真核表达载体中的TCR链高效地导入新鲜分离的SLE T细胞。研究了TCR/CD3介导的信号转导途径在X1链细胞中的恢复作用。在转TCR链的SLE T细胞中，TCR链的表面表达增加，TCR/CD3诱导的胞浆内游离钙浓度升高的反应正常，细胞底物的过度磷酸化也是如此。同时，在转染链表达载体的SLE T细胞中，Fc受体γ链的表达增加被减弱，细胞信号分子的表膜簇被重新分布到更连续的模式。TCR链置换也增加了SLE T细胞中TCR/CD3介导的IL-2的表达，并与核因子kappaB的p65亚单位在这些T细胞的核成分中的表达增加有关。这些结果提示，重建缺陷的TCR链可以逆转SLE患者T细胞中TCR/CD3介导的信号异常和IL-2的产生缺陷。
Objective. T cells from a majority of patients with systemic lupus erythematosus (SLE) display antigen receptor-mediated signaling aberrations associated with defective T cell receptor (TCR) xi chain, a subunit of the TCR/CD3 complex. This study was undertaken to explore the possibility that forced expression of TCR chain may reverse the known signaling abnormalities and defective interleukin-2 (IL-2) production in SLE T cells.Methods. Freshly isolated SLE T cells were transfected with TCR chain construct in a eukaryotic expression vector at high efficiency, by a recently developed nucleoporation technique. Restoration of TCR/ CD3-mediated signaling was studied in the xi chain-transfected cells.Results. In SLE T cells transfected with TCR chain, surface expression of TCR chain was increased and the TCR/CD3-induced increased free intracytoplasmic calcium concentration response was normalized, as was hyperphosphorylation of cellular substrates. Simultaneously, the previously noted increased expression of the Fc receptor gamma chain was diminished in SLE T cells transfected with the chain expression vector, and the surface membrane clusters of cell signaling molecules were redistributed to a more continuous pattern. TCR chain replacement also augmented the expression of diminished TCR/CD3-mediated IL-2 production in SLE T cells, associated with increased expression of the p65 subunit of nuclear factor kappaB in the nuclear fractions of these T cells.Conclusion. These results suggest that reconstitution of deficient TCR chain can reverse the TCR/ CD3-mediated signaling abnormalities as well as the defective IL-2 production in T cells of patients with SLE.