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Intracellular calcium mobilization and elastase secretion by human neutrophils in the presence of synthetic IgG Fc peptides

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6896997
Certain synthetic peptides derived from the Fc portion of human IgG were found to stimulate chemiluminescence by binding to Fc receptors for IgG and elastase secretion in human neutrophils. These peptides were tested for the ability to induce changes in intracellular Ca/sup + +/ in human neutrophils as measured by changes in fura-2/Ca/sup + +/ fluorescence. Neutrophils from normal donors were isolated using a discontinuous Percoll gradient method with a purity > 95%. Neutrophils were incubated with the acetoxymethyl ester of fura-2 (10 ..mu..M) for one hour at 37/sup 0/C. The fluorescence intensity time course at 505 nm was monitored at an excitation lambda of 340 nm. Elastase secretion was monitored by using the fluorigenic substrate MeO-suc-ala-ala-pro-val-MCA. Synthetic peptides (50 to 400..mu..g/ml) caused a dose dependent transient increase in intracellular free Ca/sup + +/ that was similar to the response by f-met-leu-phe. Phorbol 12-myristate 13-acetate (PMA) caused a decrease in fura-2/Ca/sup + +/ fluorescence. Cytochalasin B (5.0 ..mu..g/ml) was not required for the fura-2/Ca/sup + +/ response but was required for maximum elastase secretion induced by synthetic peptides, f-met-leu-phe, or PMA. These results demonstrate that stimulation of neutrophil elastase secretion by synthetic peptides is associated with changes in intracellular Ca/sup + +/.
Research Organization:
Immunetech Pharmaceuticals, San Diego, CA
OSTI ID:
6896997
Report Number(s):
CONF-8604222-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:4
Country of Publication:
United States
Language:
English