Jeron Andreas, Pfoertner Susanne, Bruder Dunja, Geffers Robert, Hammerer Peter, Hofmann Rainer, Buer Jan, Schrader Andres Jan
Department of Cell Biology, Immune Regulation Group, Helmholtz Center for Infection Research, Braunschweig, Germany.
Tumour Biol. 2009;30(3):160-70. doi: 10.1159/000228909. Epub 2009 Jul 16.
CD4(+)CD25(high) regulatory T cells (T(reg)) have the potent ability to suppress host immune responses, thus preventing autoimmune diseases. However, increased T(reg) frequencies have also been found in cancer patients implicating their involvement in tumor escape from immunological control. We investigated the frequency, functional effects and gene expression pattern of T(reg) in patients with renal cell carcinoma (RCC). Therefore, T(reg) were isolated from the peripheral blood of 11 treatment-naïve RCC patients and 11 healthy donors applying a magnetic cell separation system. Frequency, purity after isolation and function were evaluated using FACS and suppression assays, respectively. Gene expression patterns were compared applying a self-developed customized oligonucleotide microarray and by quantitative RT-PCR. T(reg) frequencies were significantly increased in RCC patients; suppression assays proved that the isolated CD4(+)CD25(high) cells had the functional characteristics of T(reg) cells. Comparing gene expression profiles between T(reg )of RCC patients and healthy controls revealed significant differences in the expression levels of 49 genes. Gene ontology identified an association of significantly up-/downregulated genes to six functional classes, particularly genes involved in apoptosis control such as LGALS1, LGALS3, BAX, IL7R and TNFRSF25. In RCC patients, frequencies of functionally active T(reg) cells were elevated; the T(reg) gene expression pattern differed significantly between patients and controls. As several anti-apoptotic genes were upregulated and pro-apoptotic genes were downregulated in RCC patients, we conclude that T(reg) cells derived from RCC patients might be less responsive to apoptotic stimuli, possibly promoting their accumulation in tumor patients.
CD4(+)CD25(high)调节性T细胞（T(reg)）具有强大的抑制宿主免疫反应的能力，从而预防自身免疫性疾病。然而，在癌症患者中也发现T(reg)频率增加，这表明它们参与了肿瘤逃避免疫控制。我们研究了肾细胞癌（RCC）患者中T(reg)的频率、功能效应和基因表达模式。因此，使用磁性细胞分离系统从11例未经治疗的RCC患者和11例健康供体的外周血中分离出T(reg)。分别使用流式细胞术（FACS）和抑制试验评估频率、分离后的纯度和功能。应用自行开发的定制寡核苷酸微阵列和定量逆转录聚合酶链反应（RT-PCR）比较基因表达模式。RCC患者的T(reg)频率显著增加；抑制试验证明分离出的CD4(+)CD25(high)细胞具有T(reg)细胞的功能特征。比较RCC患者和健康对照的T(reg)之间的基因表达谱发现49个基因的表达水平存在显著差异。基因本体论确定了显著上调/下调基因与六个功能类别的关联，特别是参与凋亡控制的基因，如半乳糖凝集素1（LGALS1）、半乳糖凝集素3（LGALS3）、 Bax蛋白、白细胞介素7受体（IL7R）和肿瘤坏死因子受体超家族成员25（TNFRSF25）。在RCC患者中，功能活跃的T(reg)细胞频率升高；患者和对照之间的T(reg)基因表达模式存在显著差异。由于RCC患者中几个抗凋亡基因上调而促凋亡基因下调，我们得出结论，源自RCC患者的T(reg)细胞可能对凋亡刺激反应较弱，这可能促进它们在肿瘤患者中的积累。
