Tsurumi H, Shimazaki M, Takahashi T, Moriwaki H, Muto Y
First Department of Internal Medicine, Gifu University School of Medicine, Japan.
Int J Hematol. 1993 Jun;57(3):213-9.
We analyzed active oxygen (hydroperoxide; H2O2) production by peripheral neutrophils in various hematological diseases by flow cytometry. One hundred microliters of heparinized fresh blood was sequentially incubated at 37 degrees C with 2',7'-dichlorofluorescein diacetate and with or without phorbol myristate acetate (PMA). After hemolysis, the pelleted white blood cells were subjected to flow cytometry, and the neutrophil fraction was gated on the cytogram. Production of H2O2 by the fraction was estimated by determining the increase in the relative intensity of fluorescence emitted from the fraction in response to stimulation by PMA. In controlled chronic myelogenous leukemia (CML) (WBC < 1 x 10(10)/1), H2O2 production was normal, while in uncontrolled CML (WBC > or = 1 x 10(10)/1), it was reduced. In myelodysplastic syndrome (MDS), H2O2 production was also reduced, but no significant difference was observed among FAB classification disease types in MDS patients. In untreated acute non-lymphocytic leukemia (ANLL), H2O2 production was reduced, while in the complete remission stage of ANLL, its level was normal, suggesting recovery from normal clones. In aplastic anemia, the H2O2 production level was normal. Steroid therapy might be responsible for the reduction of H2O2 production in non-Hodgkin's lymphoma and multiple myeloma. The production of H2O2 is closely related to the oxygen-dependent bactericidal activity of neutrophils, and, hence, can be utilized as an index to indicate susceptibility to infection. This neutrophil function can be determined easily in ordinary clinical facilities by using flow cytometry, and care should be taken to prevent infection when H2O2 production is reduced.(ABSTRACT TRUNCATED AT 250 WORDS)
我们通过流式细胞术分析了各种血液系统疾病中外周血中性粒细胞产生活性氧（过氧化氢；H2O2）的情况。将100微升肝素化新鲜血液依次在37℃下与二氯荧光素二乙酸酯以及有或无佛波酯（PMA）一起孵育。溶血后，对沉淀的白细胞进行流式细胞术检测，并在细胞图上圈出中性粒细胞部分。通过测定该部分在PMA刺激下发出的荧光相对强度的增加来估计其产生H2O2的情况。在病情得到控制的慢性粒细胞白血病（CML）（白细胞<1×10¹⁰/升）中，H2O2产生正常，而在病情未得到控制的CML（白细胞≥1×10¹⁰/升）中，其产生减少。在骨髓增生异常综合征（MDS）中，H2O2产生也减少，但MDS患者的FAB分类疾病类型之间未观察到显著差异。在未经治疗的急性非淋巴细胞白血病（ANLL）中，H2O2产生减少，而在ANLL的完全缓解期，其水平正常，提示正常克隆恢复。在再生障碍性贫血中，H2O2产生水平正常。类固醇治疗可能是导致非霍奇金淋巴瘤和多发性骨髓瘤中H2O2产生减少的原因。H2O2的产生与中性粒细胞的氧依赖性杀菌活性密切相关，因此可作为指示感染易感性的指标。通过流式细胞术在普通临床设施中可轻松测定这种中性粒细胞功能，当H2O2产生减少时应注意预防感染。（摘要截短于250字）
