Kilpinen S, Hurme M
Department of Microbiology and Immunology, University of Tampere Medical School, Finland.
Immunology. 1998 Jun;94(2):167-72. doi: 10.1046/j.1365-2567.1998.00507.x.
The capacity of neonatal T cells to secrete interleukin-2 (IL-2) has been reported to be variable. We analysed IL-2 production in purified neonatal and adult T cells using polyclonal activator phorbol ester + calcium ionophore (PDBu + iono) or receptor-mediated anti-CD3/anti-CD3+ anti-CD28 stimulation. PDBu + iono induced equally high IL-2 levels in both groups and, when stimulated with plate-bound anti-CD3 monoclonal antibody (mAb), the IL-2 secretion by neonatal cells was undetectable and adult cells produced low amounts of IL-2 (mean 331 +/- 86 pg/ml). The addition of anti-CD28 mAb to anti-CD3-stimulated cells markedly increased IL-2 production in both cell types, but levels of IL-2 in neonatal T cells remained clearly lower than those of adult T cells (respective mean values: 385 +/- 109 pg/ml and 4494 +/- 1199 pg/ml). As NF-kappa B is a critical transcription factor in the control of IL-2 expression, we next analysed its nuclear translocation in neonatal and adult T cells using the electrophoretic mobility shift assay and, because induction of reactive oxygen intermediates (ROI) is required for the activation of NF-kappa B, we also analysed levels of intracellular ROI in these cells using the ROI-reactive fluorochrome DCFH-DA and flow cytometry. In neonatal T cells NF-kappa B activation and ROI formation after anti-CD3 stimulation were low compared with adult T cells and, although addition of anti-CD28 mAb increased induction of NF-kappa B and ROI formation, levels similar to those of adults were not achieved. After PDBu + iono stimulation, the cells showed similar ROI formation and IL-2 secretion. Our results suggest that reduced IL-2 production by neonatal T cells is specific for anti-CD3 and anti-CD3+ anti-CD28-mediated stimulation and that these activators cannot effectively activate the ROI-NF-kappa B signalling pathway in neonatal T cells.
据报道，新生儿T细胞分泌白细胞介素-2（IL-2）的能力存在差异。我们使用多克隆激活剂佛波酯+钙离子载体（PDBu + iono）或受体介导的抗CD3/抗CD3 +抗CD28刺激，分析了纯化的新生儿和成人T细胞中IL-2的产生情况。PDBu + iono在两组中均诱导出同样高的IL-2水平，并且当用板结合抗CD3单克隆抗体（mAb）刺激时，新生儿细胞的IL-2分泌无法检测到，而成人细胞产生少量的IL-2（平均331±86 pg/ml）。将抗CD28 mAb添加到抗CD3刺激的细胞中，两种细胞类型的IL-2产生均显著增加，但新生儿T细胞中的IL-2水平仍明显低于成人T细胞（各自的平均值：385±109 pg/ml和4494±1199 pg/ml）。由于核因子κB（NF-κB）是控制IL-2表达的关键转录因子，我们接下来使用电泳迁移率变动分析，分析了新生儿和成人T细胞中NF-κB的核转位，并且因为NF-κB的激活需要活性氧中间体（ROI）的诱导，我们还使用ROI反应性荧光染料2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）和流式细胞术分析了这些细胞中细胞内ROI的水平。与成人T细胞相比，抗CD3刺激后新生儿T细胞中NF-κB的激活和ROI的形成较低，并且尽管添加抗CD28 mAb增加了NF-κB的诱导和ROI的形成，但未达到与成人相似的水平。在PDBu + iono刺激后，细胞显示出相似的ROI形成和IL-2分泌。我们的结果表明，新生儿T细胞中IL-2产生的减少对抗CD3和抗CD3 +抗CD28介导的刺激具有特异性，并且这些激活剂不能有效激活新生儿T细胞中的ROI-NF-κB信号通路。
