Saito T, Kuss I, Dworacki G, Gooding W, Johnson J T, Whiteside T L
University of Pittsburgh Cancer Institute, Pennsylvania 15213-2582, USA.
Clin Cancer Res. 1999 Jun;5(6):1263-73.
Proportions of apoptotic (TUNEL+) peripheral blood mononuclear cells (PBMCs) were measured by flow cytometry in patients with head and neck cancer and normal controls at the time of blood draws (0 time) and after 24-h incubation. PBMCs were incubated at 37 degrees C in medium (spontaneous apoptosis) and in the presence of CH-11 antibody (anti-Fas) or tumor necrosis factor (TNF)-alpha, both capable of inducing DNA fragmentation in activated T cells expressing the TNF family of receptors. PBMCs obtained from the patients had significantly higher (P < 0.0001) proportion of apoptotic cells than PBMCs of controls at 0 time as well as after 24-h incubation. Ex vivo apoptosis included all subsets of PBMCs: CD3+ T cells, CD16+ CD56+ natural killer cells, CD19+ B cells, and CD14+ monocytes, as determined by two-color flow cytometry. However, T cells represented the largest PBMC subset undergoing apoptosis, and lymphocytes rather than monocytes were the major TUNEL+ PBMC population. Among T cells, the level of spontaneous ex vivo apoptosis was nearly as high as that of CH-11 antibody-induced or TNF-alpha-induced apoptosis, indicating that activated Fas+ and TNFR1+ T cells were preprogrammed in vivo to die. Also, elevated levels of spontaneous apoptosis at time 0 in patients with head and neck cancer (P < 0.0001) indicated that a higher fraction of PBMCs was undergoing apoptosis in vivo in patients than controls. Together, the data suggest that an increased rate of turnover of lymphocytes is associated with cancer and may be responsible for functional lymphocyte imbalance, even in treated patients who have no evident disease.
通过流式细胞术检测头颈部癌患者和正常对照者在采血时（0 小时）及 24 小时孵育后的凋亡（TUNEL+）外周血单个核细胞（PBMC）比例。PBMC 在 37℃下于培养基中（自发凋亡）以及在 CH-11 抗体（抗 Fas）或肿瘤坏死因子（TNF）-α存在的情况下孵育，这两者都能够在表达 TNF 受体家族的活化 T 细胞中诱导 DNA 片段化。在 0 小时以及 24 小时孵育后，从患者获得的 PBMC 中凋亡细胞的比例显著高于对照组（P < 0.0001）。通过双色流式细胞术确定，体外凋亡包括 PBMC 的所有亚群：CD3+T 细胞、CD16+CD56+自然杀伤细胞、CD19+B 细胞和 CD14+单核细胞。然而，T 细胞是经历凋亡的最大 PBMC 亚群，并且淋巴细胞而非单核细胞是主要的 TUNEL+PBMC 群体。在 T 细胞中，体外自发凋亡水平几乎与 CH-11 抗体诱导或 TNF-α诱导的凋亡水平一样高，这表明活化的 Fas+和 TNFR1+T 细胞在体内预先编程死亡。此外，头颈部癌患者在 0 小时时自发凋亡水平升高（P < 0.0001），表明患者体内正在经历凋亡的 PBMC 比例高于对照组。总之，数据表明淋巴细胞周转率增加与癌症相关，并且可能是功能性淋巴细胞失衡的原因，即使在没有明显疾病的接受治疗的患者中也是如此。
