Simultaneous assay for oxidative metabolism and adhesion of human neutrophils: evidence for correlations and dissociations of the two responses

J Leukoc Biol. 1992 Apr;51(4):329-35. doi: 10.1002/jlb.51.4.329.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / pharmacology
  • Antigens, CD / physiology
  • CD11 Antigens
  • CD18 Antigens
  • Cell Adhesion / drug effects
  • Concanavalin A / administration & dosage
  • Dose-Response Relationship, Drug
  • Humans
  • In Vitro Techniques
  • N-Formylmethionine Leucyl-Phenylalanine / administration & dosage
  • Neutrophils / cytology
  • Neutrophils / metabolism
  • Neutrophils / physiology*
  • Respiratory Burst / drug effects
  • Superoxides / metabolism
  • Zymosan / administration & dosage

Substances

  • Antigens, CD
  • CD11 Antigens
  • CD18 Antigens
  • Concanavalin A
  • Superoxides
  • N-Formylmethionine Leucyl-Phenylalanine
  • Zymosan
  • Adenosine