Insoluble and soluble immune complexes activate neutrophils by distinct activation mechanisms: changes in functional responses induced by priming with cytokines.
BACKGROUND
Rheumatoid synovial fluid contains both soluble and insoluble immune complexes that can activate infiltrating immune cells such as neutrophils.


OBJECTIVE
To determine if these different complexes activate neutrophils through similar or different receptor signalling pathways. In particular, to determine the circumstances which result in the secretion of tissue damaging reactive oxygen metabolites and granule enzymes.


METHODS
Blood neutrophils were incubated with synthetic soluble and insoluble immune complexes and the ability to generate reactive oxidants tested by luminescence or spectrophotometric assays that distinguished between intracellular and extracellular production. Degranulation of myeloperoxidase and lactoferrin was determined by western blotting. The roles of FcgammaRII (CD32) and FcgammaRIIIb (CD16) were determined by incubation with Fab/F(ab')(2) fragments before activation. The effect of cytokine priming was determined by incubation with GM-CSF.


RESULTS
Insoluble immune complexes activated unprimed neutrophils, but most of the oxidants produced were intracellular. This activation required FcgammaRIIIb, but not FcgammaRII function. Soluble complexes failed to activate unprimed neutrophils but generated a rapid and extensive secretion of reactive oxygen metabolites when the cells were primed with granulocyte-macrophage colony stimulating factor (GM-CSF). This activity required both FcgammaRII and FcgammaRIIIb function. Insoluble immune complexes activated the release of granule enzymes from primed or unprimed neutrophils, but the kinetics of release did not parallel those of secretion of reactive oxygen metabolites. Only primed neutrophils released enzymes in response to soluble complexes.


CONCLUSIONS
Soluble and insoluble immune complexes activate neutrophils by separate receptor signalling pathways. Profound changes in neutrophil responsiveness to these complexes occur after cytokine priming.
Fossati
G
G
School of Biological Sciences, Life Sciences Building, University of Liverpool, Liverpool L69 7ZB, UK.
Bucknall
R C
RC
Edwards
S W
SW
D000936:Antigen-Antibody Complex / Q000502:physiology*; D015153:Blotting, Western; D002478:Cells, Cultured; D016207:Cytokines / Q000502:physiology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000502:physiology; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism; D018375:Neutrophil Activation / Q000502:physiology*; D009195:Peroxidase / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000494:pharmacology; D017452:Receptors, IgG / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology
D000936:Antigen-Antibody Complex; D016207:Cytokines; D017382:Reactive Oxygen Species; D017452:Receptors, IgG; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D009195:Peroxidase; D007781:Lactoferrin