Stimulus-dependent secretion of plasma proteins from human neutrophils.
In search for matrix proteins released from secretory vesicles of human neutrophils, a prominent 67-kD protein was identified in the extracellular medium of neutrophils stimulated by the chemotactic peptide, FMLP. The protein was purified to apparent homogeneity and partially sequenced. The sequence of the first 32 NH2-terminal amino acids was identical to the sequence of albumin. mRNA for human albumin could not be detected in bone marrow cells, nor could biosynthetic labeling of albumin be demonstrated in bone marrow cells during incubation with [14C]leucine. Immunofluorescence studies on single cells demonstrated the presence of intracellular albumin in fixed permeabilized neutrophils. Light microscopy of immunogold-silver-stained cryosections visualized albumin in cytoplasmic "granules." The morphology of these was determined by immunoelectron microscopy as vesicles of varying form and size. Subcellular fractionation studies on unstimulated neutrophils demonstrated the presence of albumin in the low density pre-gamma and gamma-regions that contain secretory vesicles, but are devoid of specific granules and azurophil granules. Albumin was readily released from these structures during activation of neutrophils with inflammatory mediators. Immunoblotting demonstrated the presence of immunoglobulin and transferrin along with albumin in exocytosed material from stimulated neutrophils. This indicates that secretory vesicles are unique endocytic vesicles that can be triggered to exocytose by inflammatory stimuli.
Borregaard
N
N
Department of Medicine and Hematology, University Hospital, Copenhagen, Denmark.
Kjeldsen
L
L
Rygaard
K
K
Bastholm
L
L
Nielsen
M H
MH
Sengeløv
H
H
Bjerrum
O W
OW
Johnsen
A H
AH
D001798:Blood Proteins / Q000378:metabolism*; D003911:Dextrans / Q000378:metabolism; D004705:Endocytosis; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism / Q000031:analogs & derivatives; D006801:Humans; D066298:In Vitro Techniques; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism* / Q000648:ultrastructure; D012709:Serum Albumin / Q000032:analysis / Q000378:metabolism
D001798:Blood Proteins; D003911:Dextrans; D012709:Serum Albumin; C015219:fluorescein isothiocyanate dextran; D009240:N-Formylmethionine Leucyl-Phenylalanine; D016650:Fluorescein-5-isothiocyanate