The effect of serine and thiol protease inhibitors on the chemiluminescence of human neutrophils in investigations in vitro.
We have studied an indirect role of serine and thiol proteases in the activation of human neutrophils in vitro. Stimulation was evaluated using a chemiluminescence (CL) generation system. Receptor-dependent and receptor-independent stimuli were studied, e.g. opsonized zymosan, formyl-methionyl-leucyl-phenylalanine, platelet activating factor, phorbol myristate acetate, and calcium ionophore A23187. The serine protease inhibitors TPCK and TLCK, and thiol protease inhibitor PHMB, diminished the CL with different potencies and in a dose-dependent manner after treatment of cells with the various stimuli. Non-specific serine protease inhibitor, PMSF, and trypsin substrate TAME, showed a low inhibitory potency with respect to CL generation. Synthetic substrates for chymotrypsin (BTEE, ATEE) significantly inhibited CL with the various stimuli used with some differences in susceptibility to their inhibition. Specific chymotrypsin inhibitors diminished both the resting and activator-induced CL. We suggest that cell-bound chymotrypsin-like protease(s) is involved in the activation of signal transduction in human neutrophils after both receptor-dependent and receptor-independent stimulation.
Kantorski
J
J
Department of Pathophysiology and Immunology, Military Medical Academy, Lódź, Poland.
Tchórzewski
H
H
D000001:Calcimycin / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014106:Tosylarginine Methyl Ester / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D015853:Cysteine Proteinase Inhibitors; D010972:Platelet Activating Factor; D015842:Serine Proteinase Inhibitors; D014107:Tosyllysine Chloromethyl Ketone; C006716:benzoyltyrosine ethyl ester; D000001:Calcimycin; D014108:Tosylphenylalanyl Chloromethyl Ketone; D014443:Tyrosine; D009240:N-Formylmethionine Leucyl-Phenylalanine; C007651:ethyl N-alpha-acetyl-tyrosinate; D014106:Tosylarginine Methyl Ester; D015054:Zymosan; D013755:Tetradecanoylphorbol Acetate