Khwaja A, Carver J E, Linch D C
Department of Haematology, University College and Middlesex School of Medicine, London, UK.
Blood. 1992 Feb 1;79(3):745-53.
Exposure of neutrophils to a range of cytokines augments their response to subsequent agonist-induced activation of the respiratory burst. We have examined the effects of several of these factors, both singly and in combination, on the priming of f-met-leu-phe (FMLP) and complement C5a-stimulated neutrophil H2O2 production, using a whole blood flow cytometric assay designed to minimize artefactual activation. Both granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF alpha) produced a similar degree of priming of the FMLP-stimulated burst in vitro (558% +/- 86%, n = 41, and 581% +/- 95%, n = 21, of the response seen with FMLP alone, respectively), but with markedly different kinetics (half-maximal response 20 minutes and 7 minutes, respectively). Preincubation with granulocyte colony-stimulating factor (G-CSF) alone caused only modest priming (202% +/- 39%, n = 14). Priming with cytokine combinations of the FMLP-stimulated burst showed that the combinations of G-CSF and TNF alpha and GM-CSF and TNF alpha are highly synergistic, with recruitment of neutrophils unresponsive to priming by single agents. Priming with the combination of GM-CSF and G-CSF was not significantly different to priming with GM-CSF alone. Similar results were obtained using C5a as the respiratory burst stimulus. Significant priming of the FMLP-stimulated respiratory burst was seen in vivo in patients receiving an infusion of GM-CSF (332% +/- 50% of preinfusion response to FMLP, P less than .005, n = 8). Priming was also seen in patients receiving G-CSF (152% +/- 58%, n = 5), although this did not reach conventional significance levels (.05 less than P less than .1). Although GM-CSF infusion caused priming in vivo, this was 48% less than predicted by preinfusion in vitro responses. This result was not due to inadequate GM-CSF levels as addition of further GM-CSF ex vivo did not correct the response. However, these neutrophils were still able to respond appropriately to ex vivo priming with TNF alpha, with a doubling in H2O2 production.
嗜中性粒细胞暴露于一系列细胞因子会增强其对随后激动剂诱导的呼吸爆发激活的反应。我们使用旨在尽量减少人为激活的全血流式细胞术检测方法，研究了其中几种因子单独及联合作用对f-甲硫氨酰-亮氨酰-苯丙氨酸（FMLP）和补体C5a刺激的嗜中性粒细胞H2O2产生的启动作用。粒细胞-巨噬细胞集落刺激因子（GM-CSF）和肿瘤坏死因子α（TNFα）在体外对FMLP刺激的爆发产生了相似程度的启动作用（分别为单独FMLP反应的558%±86%，n = 41，和581%±95%，n = 21），但动力学明显不同（半数最大反应分别为20分钟和7分钟）。单独用粒细胞集落刺激因子（G-CSF）预孵育仅引起适度的启动作用（202%±39%，n = 14）。FMLP刺激的爆发用细胞因子组合启动显示，G-CSF和TNFα以及GM-CSF和TNFα的组合具有高度协同作用，能募集对单一因子启动无反应的嗜中性粒细胞。GM-CSF和G-CSF组合启动与单独用GM-CSF启动无显著差异。以C5a作为呼吸爆发刺激物时也得到了类似结果。在接受GM-CSF输注的患者体内，FMLP刺激的呼吸爆发有显著启动作用（为输注前对FMLP反应的332%±50%，P＜0.005，n = 8）。接受G-CSF的患者也出现了启动作用（152%±58%，n = 5），尽管未达到传统的显著水平（0.05＜P＜0.1）。虽然GM-CSF输注在体内引起了启动作用，但比输注前体外反应预测的少48%。这一结果并非由于GM-CSF水平不足，因为体外添加更多GM-CSF并不能纠正反应。然而，这些嗜中性粒细胞仍能对体外TNFα启动做出适当反应，H2O2产生量增加一倍。
