Zandvliet Maarten L, Falkenburg J H Frederik, Jedema Inge, Willemze Roel, Guchelaar Henk-Jan, Meij Pauline
Department of Clinical Pharmacy and Toxicology, Leiden University Medical Center, Leiden, The Netherlands.
J Immunother. 2009 Jun;32(5):513-23. doi: 10.1097/CJI.0b013e3181a2712c.
Adoptive transfer of donor-derived cytomegalovirus (CMV)-specific T cells may provide long-lived protection from CMV disease after allogeneic stem cell transplantation. Isolation of IFNg-secreting cells after CMV peptide stimulation can be performed by IFNg capture assay to generate highly specific T-cell lines without the need for extensive culture, which may hamper their in vivo efficacy. To exploit the full potential of this approach, we analyzed the IFNg response of CMV-specific CD8+ T cells in detail. Kinetic studies showed that T-cell receptor down-regulation coincided with the induction of IFNg production upon activation, which rapidly declined thereafter despite the continued presence of specific peptide. By varying the strength of stimulation we observed that overstimulation can result in profound T-cell receptor down-regulation, more rapid decline of IFNg production and reduced expansion. On the basis of these findings, we defined optimal conditions for IFNg-based isolation of CMV-specific CD8+ T cells with maximal potential for clinical application. These data stress the importance of analyses of the kinetics of cytokine production for isolation of T cells specific for other infectious or malignant antigens to exploit the full potential of cytokine capture isolation of antigen-specific T cells.
供体来源的巨细胞病毒（CMV）特异性T细胞的过继转移可能为异基因干细胞移植后预防CMV疾病提供长期保护。在CMV肽刺激后，可通过IFNγ捕获试验分离分泌IFNγ的细胞，以生成高度特异性的T细胞系，而无需进行广泛培养，广泛培养可能会妨碍其体内疗效。为了充分发挥这种方法的潜力，我们详细分析了CMV特异性CD8+T细胞的IFNγ反应。动力学研究表明，T细胞受体下调与激活后IFNγ产生的诱导同时发生，尽管特异性肽持续存在，但此后IFNγ产生迅速下降。通过改变刺激强度，我们观察到过度刺激可导致T细胞受体深度下调、IFNγ产生更快下降以及增殖减少。基于这些发现，我们确定了基于IFNγ分离具有最大临床应用潜力的CMV特异性CD8+T细胞的最佳条件。这些数据强调了分析细胞因子产生动力学对于分离针对其他感染性或恶性抗原的特异性T细胞的重要性，以便充分发挥细胞因子捕获分离抗原特异性T细胞的潜力。
