Involvement of Fcgamma receptors and beta2 integrins in neutrophil activation by anti-proteinase-3 or anti-myeloperoxidase antibodies.
We previously described the requirement of tumour necrosis factor-alpha (TNF-alpha) and the role of beta2 integrins in the Fc-gamma receptor IIa (FcgammaRIIa)-mediated mechanism of neutrophil activation by antiproteinase-3 (anti-PR3) or anti-myeloperoxidase (anti-MPO) antibodies. In the present study, we assessed the involvement of FcgammaRIIIb by studying the respiratory burst activation of completely FcgammaRIIIb-deficient neutrophils primed by TNF-alpha and exposed to anti-PR3 or anti-MPO. Activation of the NADPH oxidase occurred normally in these neutrophils, which indicates that engagement of FcgammaRIIIb is not essential in our model. Experiments performed with neutrophils from severe leucocyte adhesion deficiency (LAD) patients confirmed that beta2 integrins play a pivotal role in this activation. We next studied whether adhesion per se, beta2-integrin-mediated adhesion, or beta2-integrin ligation without adhesion is necessary or sufficient for this activation. Anti-PR3 or anti-MPO induced an FcgammaRIIa-dependent burst in TNF-primed neutrophils incubated in wells coated with poly-L-lysine, known to induce beta2-integrin-independent adhesion, but this reaction was still inhibited by blocking CD18 antibodies. In a system with granulocyte-macrophage colony-stimulating factor (GM-CSF)-primed neutrophils, which did not enhance adhesion, we measured a similar activation by anti-PR3 or anti-MPO and inhibition by CD18. We also noticed that treatment with the beta2-integrin-activating CD18 MoAb KIM185 per se is insufficient for neutrophil activation by anti-PR3 or anti-MPO. We therefore conclude that ligation of beta2 integrins rather than adherence per se is essential for this activation, and that TNF-alpha or GM-CSF is needed for priming but not for adherence.
Reumaux
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Département d'Hématologie-Immunologie-Cytogénétique, Centre Hospitalier de Valenciennes, Valenciennes, France remaux-d@ch-valenciennes.fr
Kuijpers
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Hordijk
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PL
Duthilleul
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P
Roos
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D015703:Antigens, CD / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D018821:CD18 Antigens / Q000276:immunology*; D002448:Cell Adhesion / Q000276:immunology; D002478:Cells, Cultured; D058851:GPI-Linked Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D006801:Humans; D018370:Leukocyte-Adhesion Deficiency Syndrome / Q000276:immunology*; D053146:Myeloblastin; D018375:Neutrophil Activation / Q000276:immunology*; D009195:Peroxidase / Q000276:immunology; D017452:Receptors, IgG / Q000276:immunology*; D016897:Respiratory Burst / Q000276:immunology; D012697:Serine Endopeptidases / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D015703:Antigens, CD; D001323:Autoantibodies; D018821:CD18 Antigens; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; D017452:Receptors, IgG; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D009195:Peroxidase; D012697:Serine Endopeptidases; D053146:Myeloblastin