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  • Title: Effect of tumor necrosis factor on the generation of chlorinated oxidants by adherent human neutrophils.
    Author: Test ST.
    Journal: J Leukoc Biol; 1991 Aug; 50(2):131-9. PubMed ID: 1649239.
    Abstract:
    Human neutrophils adherent to simulated biologic surfaces undergo significant activation of the respiratory burst over prolonged periods of time in response to stimulation with the cytokines tumor necrosis factor-alpha (TNF alpha) or tumor necrosis factor-beta (TNF beta) or with the chemotactic peptide N-formyl-methionylleucylphenylalanine (FMLP). In this study, neutrophils were examined for their ability to generate the highly reactive and powerful oxidant hypochlorous acid (HOCl) and the longer-lived, less reactive endogenous nitrogen-chlorine (N-Cl) derivatives in response to these stimuli either alone or when exposed to recombinant human TNF alpha (rTNF alpha) or beta (rTNF beta) prior to addition of FMLP. Neutrophils adherent to fetal bovine serum-coated polystyrene tissue culture wells were able to generate only small quantities of HOCl when incubated with rTNF alpha, rTNF beta, or FMLP individually. However, when neutrophils were first incubated with either rTNF alpha or rTNF beta prior to addition of FMLP, there was a marked increase in HOCl generation. Neutrophils stimulated in such a manner consumed approximately 18% of the HOCl generated in the formation of N-Cl derivatives. Further scrutiny of the response to the combination of rTNF alpha and FMLP revealed that HOCl release was rapid, with 80% of total HOCl accumulation occurring within 15 min after FMLP addition. The amount of HOCl generated was dependent on the number of cells added and on the concentration of both rTNF alpha and FMLP. Comparison of HOCl generation with superoxide anion and myeloperoxidase release showed that the amount of HOCl generated was limited primarily by the amount of myeloperoxidase released rather than by the degree of respiratory burst activation. These results demonstrate that human neutrophils stimulated with FMLP after a brief incubation with rTNF alpha or rTNF beta can generate cytotoxic and microbicidal concentrations of chlorinated oxidants.
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