Interaction of human immunoglobulin G with CD16 on natural killer cells: ligand clearance, FcgammaRIIIA turnover and effects of metalloproteinases on FcgammaRIIIA-mediated binding, signal transduction and killing.
Human natural killer (NK) cells express low-affinity Fc immunoglobulin G (IgG) receptor (FcgammaRIIIA/CD16). The binding of monomeric IgG (mIgG) and F(ab')(2) fragments of 3G8 anti-CD16 monoclonal antibody (mAb) to FcgammaRIIIA was investigated by flow cytometry. Over 90% of NK cells bound endogenous IgG, and during incubation at 37 degrees C, the FcgammaRIIIA occupancy decreased slowly. Approximately 90% of NK cells bind mIgG or F(ab')(2) fragments of 3G8 anti-CD16 mAb. The calculated half-time (T(1/2)) of in vitro mIgG dissociation from FcgammaRIIIA was 130 min. By cross-linking the mIgG ligand with F(ab')(2) fragments of anti-human IgG antibody, the T(1/2) decreases to 85 min. In kinetics study, it has been shown that (125)I-mIgG bound to FcgammaRIIIA is slowly released in the culture supernatant, maybe eluted at acid pH, or partially internalized and degraded. The binding of IgG to FcgammaRIIIA was increased by 53.8% on cells cultured in the presence of RU36156, a matrix metalloproteinase (MMP) inhibitor. Furthermore, an increase in phosphorylation of Lyn tyrosine kinase, after cross-linking of mIgG-FcgammaRIIIA complex, was observed on NK cells treated with RU36156. When the FcgammaRIIIA was occupied by mIgG, the capacity of NK cells to kill K562 target cells was decreased by RU36156, because the MMP inhibitor protects CD16 from proteolysis. Our data demonstrate that binding of mIgG to human NK cells is followed by ligand dissociation and/or internalization, enzymatic degradation and exocytosis. The RU36156 MMP inhibitor protects FcgammaRIIIA from cleavage, augments NK-cell activation and may interfere in their killing capacity.
Mota
G
G
Center of Immunology, Institute of Virology, 285 Mihai Bravu st., 79650 Bucharest, Romania.
Moldovan
I
I
Calugaru
A
A
Hirt
M
M
Kozma
E
E
Galatiuc
C
C
Brasoveanu
L
L
Boltz-Nitulescu
G
G
D003601:Cytotoxicity Tests, Immunologic; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007074:Immunoglobulin G / Q000276:immunology* / Q000378:metabolism; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000276:immunology*; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000276:immunology* / Q000378:metabolism; D017452:Receptors, IgG / Q000276:immunology* / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology
D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D007074:Immunoglobulin G; D061965:Matrix Metalloproteinase Inhibitors; C106339:RU 36156; D017452:Receptors, IgG; D020782:Matrix Metalloproteinases