Lippi U, Bellavite P, Schinella M, Nicoli M, Lippi G
Laboratorio di Chimica Clinica ed Ematologia, Ospedale Civile Maggiore Piazzale Stefani 1, Verona, Italy.
Clin Lab Haematol. 1994 Mar;16(1):43-55. doi: 10.1111/j.1365-2257.1994.tb00386.x.
We have studied an alternative method to aggregometry for the assessment of human polymorphonuclear (PMN) leucocyte aggregation. This simple, rapid and reliable procedure counts unaggregated cells on both Coulter STKS and STKR haematological analysers by the impedance principle. Aggregation of PMN was induced by 15 min incubation with fresh autologous serum (FAS) after a 10 min phorbol myristate acetate (PMA) activation of neutrophils in small aliquots (0.25 ml) of suspension containing about 4.0 x 10(9) PMN/1. Differences (x 100) between count of resting and PMA+FAS treated neutrophils/count of resting PMN reflect percent aggregation. By this procedure, PMN aggregation did not occur in autologous plasma from EDTA anticoagulated whole blood; it was partially inhibited by hydrocortisone, whereas inactivated or Zymosan activated sera gave values similar to those from FAS induced aggregation. PMA aggregation was dependent on Ca2+ + Mg2+ concentration. Intra-assay analytical variability did not exceed 4% on either instrument. Reference values (n = 20) of percent PMN aggregation were 50.7 +/- 4.7 on STKS and 47.1 +/- 4.8 on STKR. Most probably, the interindividual variance was due to the physiological variability of Mg2+ and/or Ca2+ concentrations in FAS. Thus, this procedure reflects the true PMN aggregability status in a given subject, and in a given electrolyte environment.
我们研究了一种用于评估人类多形核(PMN)白细胞聚集的凝集测定替代方法。这种简单、快速且可靠的程序通过阻抗原理在库尔特STKS和STKR血液分析仪上对未聚集的细胞进行计数。在含有约4.0×10⁹个PMN/1的小份(0.25 ml)悬浮液中,嗜中性粒细胞经佛波酯肉豆蔻酸酯乙酸酯(PMA)激活10分钟后,与新鲜自体血清(FAS)孵育15分钟,诱导PMN聚集。静息中性粒细胞计数与经PMA + FAS处理的中性粒细胞计数/静息PMN计数之间的差异(×100)反映聚集百分比。通过该程序,在来自EDTA抗凝全血的自体血浆中未发生PMN聚集;它被氢化可的松部分抑制,而热灭活或酵母聚糖激活的血清给出的值与FAS诱导聚集的值相似。PMA聚集取决于Ca²⁺ + Mg²⁺浓度。两种仪器的批内分析变异性均不超过4%。在STKS上PMN聚集百分比的参考值(n = 20)为50.7±4.7,在STKR上为47.1±4.8。个体间差异很可能是由于FAS中Mg²⁺和/或Ca²⁺浓度的生理变异性所致。因此,该程序反映了给定个体在给定电解质环境中的真实PMN聚集能力状态。