Thomas Alissa A, Fisher Jan L, Rahme Gilbert J, Hampton Thomas H, Baron Udo, Olek Sven, Schwachula Tim, Rhodes C Harker, Gui Jiang, Tafe Laura J, Tsongalis Gregory J, Lefferts Joel A, Wishart Heather, Kleen Jonathan, Miller Michael, Whipple Chery A, de Abreu Francine B, Ernstoff Marc S, Fadul Camilo E
Department of Neurology, Dartmouth Hitchcock Medical Center, Lebanon, New Hampshire (A.A.T.); Department of Medicine, Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire (J.L.F.); Department of Genetics, Geisel School of Medicine at Dartmouth, Norris Cotton Cancer Center, Dartmouth College, Lebanon, New Hampshire (G.J.R.); Epiontis GmbH, Berlin, Germany (U.B., S.O., T.S.); Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, New Hampshire (C.H.R., L.J.T., G.J.T., J.A.L., F.B.d.A.); Section of Biostatistics and Epidemiology, Department of Family and Community Medicine, Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire (J.G.); Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire (T.H.H.); Department of Psychiatry, Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire (H.W.); Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire (J.K., M.M.); Department of Medicine, Norris Cotton Cancer Center, Geisel School of Medicine, Dartmouth Hitchcock Medical Center, Lebanon, New Hampshire (C.A.W.); Melanoma Program, Taussig Cancer Institute, Cleveland Clinic Foundation, Cleveland, Ohio (M.S.E.); Department of Medicine, Norris Cotton Cancer Center, Geisel School of Medicine at Dartmouth, Dartmouth Hitchcock Medical Center, Lebanon, New Hampshire (C.E.F.).
Neuro Oncol. 2015 Jun;17(6):801-9. doi: 10.1093/neuonc/nou363. Epub 2015 Jan 24.
Regulatory T cells (Tregs) are potentially prognostic indicators in patients with glioblastoma. If differences in frequency of Tregs in tumor or blood account for substantial variation in patient survival, then reliably measuring Tregs may enhance treatment selection and improve outcomes.
We measured Tregs and CD3+ T cells in tumors and blood from 25 patients with newly diagnosed glioblastoma. Tumor-infiltrating Tregs and CD3+ T cells, measured by quantitative DNA demethylation analysis (epigenetic qPCR) and by immunohistochemistry, and peripheral blood Treg proportions measured by flow cytometry were correlated with patient survival. Additionally, we analyzed data from The Cancer Genome Atlas (TCGA) to correlate the expression of Treg markers with patient survival and glioblastoma subtypes.
Tregs, as measured in tumor tissue and peripheral blood, did not correlate with patient survival. Although there was a correlation between tumor-infiltrating Tregs expression by epigenetic qPCR and immunohistochemistry, epigenetic qPCR was more sensitive and specific. Using data from TCGA, mRNA expression of Forkhead box protein 3 (FoxP3) and Helios and FoxP3 methylation level did not predict survival. While the classical glioblastoma subtype corresponded to lower expression of Treg markers, these markers did not predict survival in any of the glioblastoma subtypes.
Although immunosuppression is a hallmark of glioblastoma, Tregs as measured in tissue by gene expression, immunohistochemistry, or demethylation and Tregs in peripheral blood measured by flow cytometry do not predict survival of patients. Quantitative DNA demethylation analysis provides an objective, sensitive, and specific way of identifying Tregs and CD3+ T cells in glioblastoma.
调节性T细胞（Tregs）可能是胶质母细胞瘤患者的预后指标。如果肿瘤或血液中Tregs频率的差异导致患者生存率出现显著差异，那么可靠地检测Tregs可能会优化治疗选择并改善治疗结果。
我们检测了25例新诊断胶质母细胞瘤患者肿瘤组织和血液中的Tregs及CD3+ T细胞。通过定量DNA去甲基化分析（表观遗传定量聚合酶链反应）和免疫组织化学检测肿瘤浸润性Tregs及CD3+ T细胞，通过流式细胞术检测外周血Treg比例，并将其与患者生存率进行关联分析。此外，我们分析了癌症基因组图谱（TCGA）的数据，以将Treg标志物的表达与患者生存率及胶质母细胞瘤亚型进行关联。
肿瘤组织和外周血中检测到的Tregs与患者生存率无相关性。尽管通过表观遗传定量聚合酶链反应和免疫组织化学检测的肿瘤浸润性Tregs表达之间存在相关性，但表观遗传定量聚合酶链反应更敏感且特异。利用TCGA的数据，叉头框蛋白3（FoxP3）、Helios的mRNA表达及FoxP3甲基化水平均不能预测生存率。虽然经典型胶质母细胞瘤亚型对应较低的Treg标志物表达，但这些标志物在任何胶质母细胞瘤亚型中均不能预测生存率。
尽管免疫抑制是胶质母细胞瘤的一个特征，但通过基因表达、免疫组织化学或去甲基化在组织中检测到的Tregs以及通过流式细胞术在外周血中检测到的Tregs均不能预测患者的生存率。定量DNA去甲基化分析为识别胶质母细胞瘤中的Tregs及CD3+ T细胞提供了一种客观、敏感且特异的方法。
