Further characterization of the NB 1 antigen as a variably expressed 56-62 kD GPI-linked glycoprotein of plasma membranes and specific granules of neutrophils.
The human neutrophil-specific alloantigen NB1 was identified as a glycosyl-phosphatidylinositol (GPI)-anchored N-glycosylated protein of M(r) 56-62 kD under reducing conditions. Under non-reducing conditions its M(r) was 49-55 kD. This glycoprotein antigen was found to be expressed by only a subpopulation of normal donor neutrophils, and could not be detected on other blood cells. The allotypic epitope recognized by human anti-NB1 IgG was also recognized by the mouse monoclonal antibody 1B5. The percentage of neutrophils stained by these antibodies varied greatly among healthy donors (range 0-100%). When 16 donors were repeatedly tested, the NB1-positive neutrophil fraction appeared to remain remarkably constant over time in most donors, but significant fluctuations were seen in some. NB1 antigen was found to be expressed not only on the plasma membrane, but also intracellularly on the membranes of small vesicles and specific granules. The neutrophils which expressed NB1 antigen on the plasma membrane were the same as those with intracellular expression of this antigen. Crosslinking of NB1 antigen on the plasma membrane with monoclonal antibody 1B5 and goat-anti-mouse Ig resulted in internalization of the complex, while in-vitro stimulation of neutrophils caused an increase of the intensity of plasma membrane staining with anti-NB1, but only of those cells that were positive already prior to stimulation. The NB1 glycoprotein thus appears to identify a distinct subset of neutrophils, the size of which greatly varies among healthy donors. The function of the NB1 glycoprotein remains unclear, but its behaviour upon crosslinking and chemotactic peptide stimulation suggests a possible role as receptor molecule.
Goldschmeding
R
R
Department of Haematology, The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
van Dalen
C M
CM
Faber
N
N
Calafat
J
J
Huizinga
T W
TW
van der Schoot
C E
CE
Clement
L T
LT
von dem Borne
A E
AE
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002462:Cell Membrane / Q000276:immunology / Q000648:ultrastructure; D003594:Cytoplasmic Granules / Q000276:immunology* / Q000648:ultrastructure; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D058851:GPI-Linked Proteins; D006801:Humans; D007519:Isoantigens / Q000097:blood*; D008562:Membrane Glycoproteins / Q000097:blood*; D016253:Microscopy, Immunoelectron; D009504:Neutrophils / Q000276:immunology* / Q000648:ultrastructure; D011233:Precipitin Tests; D011956:Receptors, Cell Surface
D000911:Antibodies, Monoclonal; C407242:CD177 protein, human; D000939:Epitopes; D058851:GPI-Linked Proteins; D007519:Isoantigens; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface