Rothe G, Kellermann W, Valet G
Mildred-Scheel-Labor für Krebszellforschung, Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
J Lab Clin Med. 1990 Jan;115(1):52-61.
Flow cytometric parameters of neutrophil function, such as phagocytosis and degradation of Escherichia coli, intracellular pH value, esterase activity, and cell volume, were evaluated as risk indicators for sepsis- and trauma-related pulmonary and cardiovascular organ failure in intensive care patients. Serial blood samples (n = 201) were obtained from 47 prospectively identified patients. Each patient's condition was classified daily within four categories: post-traumatic (n = 22) or septic (n = 28) organ failure, transition state (n = 119), and stable organ function after recovery (n = 27). Thirty-two parameters of neutrophil function were automatically calculated for each blood sample from several flow cytometric list mode measurements of cell samples vitally stained with acridine orange for intact and denatured DNA or with 1,4-diacetoxy-2,3-dicyanobenzene for intracellular pH and esterase activity. The DNA of dead cells was simultaneously counterstained with propidium iodide. The cell biochemical parameter pattern was significantly different among samples of patients from the four clinical categories (p less than 0.05). Hyperergic phagocytosis was observed after trauma, in contrast to hypoergic phagocytosis, increased neutrophil cell volume, and elevated intracellular pH during sepsis. The clinical categories were correctly identified in 82% of the samples by automated classification with the DIAGNOS1/SPSS program system from the flow cytometrically determined cell functions. The course of the disease was correctly predicted 3 days in advance to the clinical manifestation of pulmonary or cardiovascular organ failure in 92% of the samples. The multifunctional analysis of neutrophils by flow cytometry seems of interest for early medical intervention in preseptic and preshock patients.
中性粒细胞功能的流式细胞术参数,如对大肠杆菌的吞噬作用和降解、细胞内pH值、酯酶活性和细胞体积,被评估为重症监护患者败血症和创伤相关肺及心血管器官衰竭的风险指标。从47例前瞻性确定的患者中采集了系列血样(n = 201)。每天将每位患者的病情分为四类:创伤后(n = 22)或败血症性(n = 28)器官衰竭、过渡状态(n = 119)以及恢复后器官功能稳定(n = 27)。通过对用吖啶橙对完整和变性DNA进行活体染色或用1,4 - 二乙酰氧基 - 2,3 - 二氰基苯对细胞内pH和酯酶活性进行活体染色的细胞样本进行多次流式细胞术列表模式测量,为每个血样自动计算32个中性粒细胞功能参数。死细胞的DNA同时用碘化丙啶复染。来自四个临床类别的患者样本中细胞生化参数模式有显著差异(p小于0.05)。与低反应性吞噬作用相反,创伤后观察到高反应性吞噬作用,败血症期间中性粒细胞细胞体积增加且细胞内pH升高。使用来自流式细胞术测定的细胞功能的DIAGNOS1/SPSS程序系统进行自动分类,在82%的样本中正确识别了临床类别。在92%的样本中,在肺或心血管器官衰竭临床表现前3天就正确预测了疾病进程。流式细胞术对中性粒细胞的多功能分析似乎对败血症前期和休克前期患者的早期医学干预具有意义。