Logan T F, Gooding W, Kirkwood J M, Shadduck R K
University of Pittsburgh, PA, USA.
Exp Hematol. 1996 Jan;24(1):49-53.
In humans, tumor necrosis factor (TNF) treatment has been associated with characteristic changes in circulating white blood cell populations (leukopenia followed by leukocytosis) and increased cell-surface expression of integrins. A similar pattern of effects on leukocytes occurs with granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF treatment. To determine whether these effects were caused directly by TNF or as a result of secondary CSF release, G-GM-, and M-CSF levels were measured after TNF infusion (9.6 x 10(6) U/mg protein; < 5.0 endotoxin U/mg protein) in cancer patients during two phase I trials of TNF. One patient with aggressive fibromatosis was treated with TNF alone (200 micrograms/m2, days 1-5 every third week) and 10 patients (four colon cancer, four head and neck cancer; one melanoma; one sarcoma) received mitomycin C (15 mg/m2, day 1) followed by TNF (60-180 micrograms/m2, days 1-3) every sixth week. All treatments were given IV, mitomycin C over 5 minutes and TNF over 2 hours. Serum samples were collected at times 0 (before mitomycin C and TNF) and 1, 2, 4, 6, 12, and 24 hours after TNF initiation on day 1 and at similar times on subsequent treatment days. M-CSF samples were analyzed by radioimmunoassay (RIA) and G-CSF and GM-CSF by ELISA. The mean baseline M-CSF levels in normal control subjects (n = 12) was 158.4 +/- 36.2 (SD) U/mL, and in pretreatment cancer patients (n = 10) 235.7 +/- 60.9 U/mL (p = 0.004, Wilcoxon test). M-CSF levels increased 4 hours after TNF initiation (mean 354.7 +/- 96.3 U/mL; p = 0.020), remained elevated at 6 hours (305.6 +/- 45.4 U/mL; p = 0.004, Wilcoxon signed-rank test), and subsequently declined. This pattern was seen in all patients treated with TNF, whether treatment was TNF alone or TNF with mitomycin C. In patients treated with mitomycin C and TNF, G-CSF levels increased at 4 hours after TNF initiation (mean 3886 +/- 2009 pg/mL; p = 0.004), remained elevated at 6 hours (mean 2140 +/- 1131 pg/mL; p = 0.004), and subsequently declined. GM-CSF levels were not measurable before or after treatment with TNF. The changes in all three endogenous cytokines were not temporally related to the previously described leukopenia and integrin upregulation on circulating leukocytes and, therefore, appear to be unrelated to this event. However, release of endogenous G-CSF and M-CSF under the influence of TNF does temporally coincide with the previously described leukocytosis, suggesting a possible role for these endogenous cytokines in the release of bone marrow cellular stores.
在人类中，肿瘤坏死因子（TNF）治疗与循环白细胞群体的特征性变化（白细胞减少后白细胞增多）以及整合素细胞表面表达增加有关。粒细胞 - 巨噬细胞集落刺激因子（GM - CSF）和粒细胞集落刺激因子（G - CSF）治疗对白细胞也会产生类似的效应模式。为了确定这些效应是由TNF直接引起的，还是继发于脑脊液释放的结果，在两项TNF的I期临床试验中，对癌症患者输注TNF（9.6×10⁶ U/mg蛋白质；<5.0内毒素U/mg蛋白质）后，测量了G - CSF、GM - CSF和M - CSF的水平。一名侵袭性纤维瘤病患者单独接受TNF治疗（200微克/m²，第1 - 5天，每三周一次），10名患者（4例结肠癌、4例头颈癌、1例黑色素瘤、1例肉瘤）先接受丝裂霉素C（15 mg/m²，第1天）治疗，然后每六周接受TNF（60 - 180微克/m²，第1 - 3天）治疗。所有治疗均通过静脉注射给药，丝裂霉素C注射5分钟，TNF注射2小时。在第1天TNF开始注射前的0时（在丝裂霉素C和TNF之前）以及注射后1、2、4、6、12和24小时采集血清样本，并在后续治疗日的类似时间采集样本。M - CSF样本通过放射免疫测定法（RIA）分析，G - CSF和GM - CSF样本通过酶联免疫吸附测定法（ELISA）分析。正常对照受试者（n = 12）的平均基线M - CSF水平为158.4±36.2（标准差）U/mL，预处理癌症患者（n = 10）的平均基线M - CSF水平为235.7±60.9 U/mL（p = 0.004，Wilcoxon检验）。TNF开始注射后4小时M - CSF水平升高（平均354.7±96.3 U/mL；p = 0.020），6小时时仍保持升高（305.6±45.4 U/mL；p = 0.004，Wilcoxon符号秩检验），随后下降。在所有接受TNF治疗的患者中均观察到这种模式，无论治疗是单独使用TNF还是TNF联合丝裂霉素C。在接受丝裂霉素C和TNF治疗的患者中，TNF开始注射后4小时G - CSF水平升高（平均3886±2009 pg/mL；p = 0.004），6小时时仍保持升高（平均2140±1131 pg/mL；p = 0.004），随后下降。在TNF治疗前后均无法检测到GM - CSF水平。所有三种内源性细胞因子的变化在时间上与先前描述的循环白细胞白细胞减少和整合素上调无关，因此，似乎与该事件无关。然而，在TNF影响下内源性G - CSF和M - CSF的释放确实与先前描述的白细胞增多在时间上一致，提示这些内源性细胞因子在骨髓细胞储存释放中可能发挥作用。
