Strauss Laura, Bergmann Christoph, Whiteside Theresa L
Department of Pathology, University of Pittsburgh School of Medicine and Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Int J Cancer. 2007 Dec 1;121(11):2473-83. doi: 10.1002/ijc.23001.
Circulating human CD4(+)CD25(high)Foxp3(+) T cell populations (Treg) may contain activated CD4(+)CD25(+) T cells interfering with Treg evaluation. To gain insights into the phenotypic and functional characteristics of Treg in patients with cancer, we have analyzed CD4(+)CD25(high) populations at the clonal level. Single-cell sorted (SCS) CD4(+)CD25(high) T cells obtained from PBMC of normal controls (NC) or patients with squamous cell carcinoma of the head and neck (HNSCC) were plated at 1 cell/well in 96 well plates and expanded with anti-CD3/anti-CD28 Abs and 1,000 IU IL-2/mL in the presence or absence of rapamycin (1 nM). All generated clones were evaluated for the phenotype by flow cyometry and suppressor function in CFSE-based proliferation assays. Clones had heterogeneous CD25 expression levels. Cloning efficiency of CD4(+)CD25(high) T cells was low. CD25(high) clones expressed CTLA-4, Foxp3, CD62L, but little GITR and suppressed proliferation of autologous CD4(+)CD25(-) responder cells. Clones of activated CD4(+)CD25(interm./low) cells expressed intermediate to high levels of GITR and HLA-DR and did not suppress proliferation of responder cells. The number, suppressor phenotype and function of CD25(high) Treg clones were significantly enhanced in HNSCC patients relative to NC (p </= 0.001). CD4(+)CD25(+) populations comprise phenotypically and functionally distinct subsets of CD25(+) cells. Only a small fraction of these activated CD4(+) T cells are potent suppressor cells characterized by high expression levels of CD25, Foxp3, CTLA-4 and CD62L. The number of expandable Treg is increased in HNSCC patients.
循环中的人CD4(+)CD25(高)Foxp3(+) T细胞群体（调节性T细胞）可能包含干扰调节性T细胞评估的活化CD4(+)CD25(+) T细胞。为深入了解癌症患者中调节性T细胞的表型和功能特征，我们在克隆水平分析了CD4(+)CD25(高)群体。从正常对照（NC）或头颈部鳞状细胞癌（HNSCC）患者的外周血单核细胞（PBMC）中获得的单细胞分选（SCS）CD4(+)CD25(高) T细胞，以1个细胞/孔接种于96孔板中，并在有或无雷帕霉素（1 nM）存在的情况下，用抗CD3/抗CD28抗体和1000 IU IL-2/mL进行扩增。通过流式细胞术评估所有产生的克隆的表型，并在基于羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）的增殖试验中评估其抑制功能。克隆具有异质性的CD25表达水平。CD4(+)CD25(高) T细胞的克隆效率较低。CD25(高)克隆表达细胞毒性T淋巴细胞相关抗原4（CTLA-4）、叉头框蛋白3（Foxp3）、淋巴细胞功能相关抗原1（CD62L），但几乎不表达糖皮质激素诱导的肿瘤坏死因子受体（GITR），并抑制自体CD4(+)CD25(-)反应细胞的增殖。活化的CD4(+)CD25(中等/低)细胞的克隆表达中等至高水平的GITR和人白细胞抗原DR（HLA-DR），且不抑制反应细胞的增殖。相对于NC，HNSCC患者中CD25(高)调节性T细胞克隆的数量、抑制表型和功能显著增强（p≤0.001）。CD4(+)CD25(+)群体包含表型和功能不同的CD25(+)细胞亚群。这些活化的CD4(+) T细胞中只有一小部分是强效抑制细胞，其特征为CD25、Foxp3、CTLA-4和CD62L的高表达水平。HNSCC患者中可扩增的调节性T细胞数量增加。
