SIMULTANEOUS ASSAY FOR OXIDATIVE-METABOLISM AND ADHESION OF HUMAN NEUTROPHILS - EVIDENCE FOR CORRELATIONS AND DISSOCIATIONS OF THE 2 RESPONSES

被引:50
作者
BELLAVITE, P
CHIRUMBOLO, S
MANSOLDO, C
GANDINI, G
DRI, P
机构
[1] UNIV VERONA, IST CHIM & MICROSCOPIA CLIN, I-37100 VERONA, ITALY
[2] UNIV TRIESTE, IST PATOL GEN, I-34127 TRIESTE, ITALY
[3] OSPED POLICLIN VERONA, CTR IMMUNOTRASFUS, VERONA, ITALY
关键词
ADHESION; SUPEROXIDE ANION; NEUTROPHIL ACTIVATION; NEUTROPHIL INHIBITORS; PHAGOCYTOSIS;
D O I
10.1002/jlb.51.4.329
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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.
引用
收藏
页码:329 / 335
页数:7
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