Fletcher M P, Vassar M J, Holcroft J W
Department of Internal Medicine, University of California, Davis 95616.
Inflammation. 1988 Oct;12(5):455-73. doi: 10.1007/BF00919439.
Neutrophils (PMNs) from patients with adult respiratory distress syndrome (ARDS) were assessed for light scattering, membrane potential, and phagocytic responses using fluorescent probes and flow cytometry to evaluate individual cells. Qualitative and quantitative oxidant responses were measured by nitroblue tetrazolium (NBT) and cytochrome c reduction assays, respectively. The results were correlated with the proportion of cells binding the PMN subset-specific monoclonal antibody 31D8. Despite an increased forward scatter signal (4.3 +/- 1.6 vs. 1.3 +/- 1.1 ARDS vs. control, P = 0.041) and spontaneous NBT test (12.6 +/- 4.7% vs. 2.5 +/- 0.8% positive, ARDS vs. control, P = 0.033) indicating in vivo priming of ARDS PMNs, there were no significant differences between ARDS and control PMNs in assays of stimulated membrane potential, NBT, and O.2- production or phagocytosis. However, positive correlations between the degree of prestimulus forward light scatter and subsequent O.2- production to FMLP (r = 0.673, P = 0.006) and between the percentage of bands and the O.2- response to PMA (r = 0.660, P = 0.003), suggest that the great variability of the ARDS PMN functional responses may relate to varying degrees of in vivo cell priming and/or deactivation. ARDS PMNs demonstrated a significantly lower percentage of 31D8 positive cells (73.4 +/- 7.5% vs. 94.5 +/- 1.6%, P = 0.012) and a lower level of 31D8 staining when compared to normals (60.1 +/- 10.4% of control level, P = 0.001). The lower 31D8 expression did not directly correlate with any functional parameter tested or with the proportion of immature cells. However, patients receiving an intravenous PGE1 infusion demonstrated a significant increase in 31D8 staining relative to controls and inhibition of PMA-stimulated O.2- production. The data suggest that the function of PMNs from ARDS patients varies widely and reflects great in vivo variation in cell priming. While the mechanism responsible for the lowered expression of the 31D8 antigen and its apparent modulation by PGE1 is unknown, 31D8 may be an indirect marker for in vivo stress factors that regulate the preferential release of a structurally distinct PMN subset from the bone marrow.
利用荧光探针和流式细胞术评估成人呼吸窘迫综合征（ARDS）患者的中性粒细胞（PMNs）的光散射、膜电位和吞噬反应，以评估单个细胞。分别通过硝基蓝四氮唑（NBT）和细胞色素c还原试验测定定性和定量的氧化剂反应。结果与结合PMN亚群特异性单克隆抗体31D8的细胞比例相关。尽管前向散射信号增加（ARDS组为4.3±1.6，对照组为1.3±1.1，P = 0.041）以及自发NBT试验阳性（ARDS组为12.6±4.7%，对照组为2.5±0.8%，P = 0.033）表明ARDS患者的PMN在体内被激活，但在刺激膜电位、NBT和超氧阴离子（O₂⁻）产生或吞噬作用测定中，ARDS组和对照组的PMN之间没有显著差异。然而，刺激前前向光散射程度与随后对甲酰甲硫氨酸-亮氨酸-苯丙氨酸（FMLP）的O₂⁻产生之间呈正相关（r = 0.673，P = 0.006），以及带状细胞百分比与对佛波酯（PMA）的O₂⁻反应之间呈正相关（r = 0.660，P = 0.003），这表明ARDS患者PMN功能反应的巨大变异性可能与体内细胞激活和/或失活的不同程度有关。与正常组相比，ARDS患者的PMN显示31D8阳性细胞百分比显著降低（73.4±7.5%对94.5±1.6%，P = 0.012），31D8染色水平也较低（为对照组水平的60.1±10.4%，P = 0.001）。较低的31D8表达与所测试的任何功能参数或未成熟细胞比例均无直接相关性。然而，接受静脉注射前列腺素E1（PGE1）的患者相对于对照组显示31D8染色显著增加，且PMA刺激的O₂⁻产生受到抑制。数据表明，ARDS患者的PMN功能差异很大，反映出体内细胞激活存在很大差异。虽然导致31D8抗原表达降低及其被PGE1明显调节的机制尚不清楚，但31D8可能是体内应激因素的间接标志物，这些应激因素调节骨髓中结构不同的PMN亚群的优先释放。
