Reversible blockade of the respiratory burst in human neutrophils by a cleavable cross-linking agent.
Human neutrophils treated with low concentrations of the homobifunctional cross-linking reagents disuccinimidyl suberate and dithiobis(succinimidylpropionate) failed to generate superoxide in response to any of several stimuli, including phorbol myristate acetate, fMet-Leu-Phe, A23187, fluoride, and opsonized zymosan. The cross-linking reagent interfered with the activation of NADPH oxidase, but not with its activity. Cells treated with succinimidyl butyrate, a monovalent analog of the cross-linkers, underwent a normal respiratory burst. Cross-linking also inhibited degranulation, phagocytosis, and fluorescence responses of potential-sensitive dyes but had little effect on lactate production, sugar transport, the binding of fMet-Leu-Phe, or the activity of various enzymes in the cross-linked neutrophils. Most of the cellular functions inhibited through the reaction of neutrophils with the cleavable cross-linker dithiobis(succinimidylpropionate) could be restored by reduction of the disulfide bonds of the cell-bound cross-linker with dithiothreitol.
D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D006801:Humans; D009113:Muramidase / Q000097:blood; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009247:NADH, NADPH Oxidoreductases / Q000097:blood; D019255:NADPH Oxidases; D009504:Neutrophils / Q000187:drug effects*; D010101:Oxygen Consumption / Q000187:drug effects*; D013388:Succinimides / Q000494:pharmacology*; D013481:Superoxides / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D003432:Cross-Linking Reagents; D013388:Succinimides; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D009113:Muramidase; C011240:dithiobis(succinimidylpropionate); D013755:Tetradecanoylphorbol Acetate; D004229:Dithiothreitol; C019358:disuccinimidyl suberate