SIMULTANEOUS ASSAY FOR OXIDATIVE-METABOLISM AND ADHESION OF HUMAN NEUTROPHILS - EVIDENCE FOR CORRELATIONS AND DISSOCIATIONS OF THE 2 RESPONSES
SIMULTANEOUS ASSAY FOR OXIDATIVE-METABOLISM AND ADHESION OF HUMAN NEUTROPHILS - EVIDENCE FOR CORRELATIONS AND DISSOCIATIONS OF THE 2 RESPONSES
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DOI：
10.1002/jlb.51.4.329
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发表时间：
1992-04-01
影响因子：
5.5
通讯作者：
DRI, P
中科院分区：
文献类型：
--
作者：
BELLAVITE, P;CHIRUMBOLO, S;DRI, P
描述了一种同时评估人类中性粒细胞氧化代谢和粘附性的测定方法，以及某些特定的应用。在血清包被的微滴板中进行孵育，氧化代谢以O2释放来测量，洗掉非粘附细胞后，以酸性磷酸酶活性来测量粘附。在该系统中，三种激动剂——豆豆蛋白A和n -甲酰基-蛋氨酸-亮基-苯丙氨酸——都能诱导O2释放和细胞粘附的激活，但这两种功能具有时间过程和剂量依赖模式，取决于刺激物。特别是刀豆蛋白A， O2释放和粘附反应明显分离；这种凝集素在低剂量下增加了中性粒细胞的粘附性而不触发任何O2的产生，而在高剂量下它既增加了O2的产生又增加了粘附性。抗整合素单克隆抗体对低剂量刀豆蛋白A诱导的粘附无影响，但对其他受体激动剂诱导的粘附有抑制作用。研究还发现，NADPH氧化酶抑制剂二苯碘、巯基试剂n -乙基马来酰亚胺和A2激动剂腺苷对粘附和O2产生的影响也不同，这表明这些中性粒细胞反应具有不同的转导途径，也取决于刺激的类型。
An assay method for the simultaneous evaluation of the oxidative metabolism and adherence of human neutrophils is described, together with certain specific applications. Incubations were performed in serum-coated microtiter plates, where oxidative metabolism was measured as O2- release and, after washing out the nonadherent cells, the adhesion was measured as activity of acid phosphatase. Three agonists tested in this system-opsonized zymosan, concanavalin A, and N-formyl-methionyl-leucyl-phenylalanine - induced both activation of O2- release and cell adhesion, but the two functions had time course and dose dependence patterns that varied depending on the stimulant. Particularly with concanavalin A, O2- release and adhesion response were markedly dissociated; this lectin at low doses increased neutrophil adherence without triggering any O2- production, whereas at high doses it increased both O2- production and adherence. Anti-integrin monoclonal antibodies did not affect adhesion induced by low-dose concanavalin A but inhibited the adhesion induced by the other tested agonists. Adhesion and O2- production were also found to be differentially affected by the NADPH oxidase inhibitor diphenylene iodonium, the sulfhydryl reagent N-ethylmaleimide and the A2 agonist adenosine, indicating that these neutrophil responses have various transductional pathways that also depend on the type of stimulus.