Enhancement by growth hormone of phorbol diester-stimulated respiratory burst in human polymorphonuclear leukocytes.
The oxidative metabolic burst of mononuclear and polymorphonuclear phagocytes can be stimulated to produce free oxygen radicals. Several substances can enhance this respiratory burst activity by a priming action: recently growth hormone (rat and porcine) was demonstrated to act as a priming agent on rat peritoneal and on porcine alveolar macrophages. In our study we wanted to verify whether also human GH had a similar priming action on homologous cells, in particular on polymorphonuclear leukocytes. To determine the oxidative activity of polymorphonuclear leukocytes, after stimulation with phorbol myristate acetate, a flow-cytometric assay was employed which registered the intracellular formation of highly fluorescent products as indicators for the intracellular formation of hydrogen peroxide. The incubation of phorbol myristate acetate-stimulated polymorphonuclear leukocytes with GH resulted in a time-dependent and dose-dependent increase in fluorescence, thus demonstrating that human GH enhances in vitro the oxidative metabolic burst of these cells. The action of GH appeared to be significant after 30 min of incubation, was maximal at 60 min, and decreased after 90 min. After one hour of incubation, the first significant variation of fluorescence appeared with GH at a concentration of 50 micrograms/l. The maximum effect was seen at 100 micrograms/l with no further increase. Specificity of GH action was demonstrated by the inhibition of its effect by the addition of monoclonal antibodies to GH.
Spadoni
G L
GL
Department of Pediatrics, 2nd University of Rome, Italy.
Spagnoli
A
A
Cianfarani
S
S
Del Principe
D
D
Menichelli
A
A
Di Giulio
S
S
Boscherini
B
B
D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000502:physiology; D015415:Biomarkers / Q000097:blood; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins; D005453:Fluorescence; D013006:Growth Hormone / Q000276:immunology / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D010101:Oxygen Consumption / Q000187:drug effects*; D010703:Phorbol Esters / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005452:Fluoresceins; D007328:Insulin; D010703:Phorbol Esters; C037631:2',7'-dichlorofluorescein; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone; D006861:Hydrogen Peroxide; D013755:Tetradecanoylphorbol Acetate