Antinucleosome autoantibodies bind directly to cell lines in vitro and via the FcgammaRIIB receptor to B lymphocytes in vivo: a role for immune complexes in interactions between antinucleosome IgG2a and B cells of BXSB lupus mice.
The initial novel observation of this study was that most B cells of male BXSB lupus mice bear surface IgG2a(b) of extrinsic origin. To define the surface antigen, we here examine three (NZBxBXSB)F1-derived IgG2a(b) monoclonal antibodies (mAbs) selected for binding to cell surfaces. Surprisingly, all three mAbs bound the nucleosome (nuc) particle, the fundamental unit of chromatin and an early target of autoimmunity in systemic lupus erythematosus. Their tentative dissociation constant (K(d)) for soluble nuc particles was approximately 7 x 10(-10) m. The mAbs bound more weakly to both H2A-H2B-DNA and H3-H4-DNA complexes, and in immunoblot they stained all four core histones. The mAbs detected a surface antigen on all cell lines examined, present on viable cells. When stripped of nuc, and in the presence of DNase I, their binding to cell lines improved. Heparin displaced the antigen from the cell surface. In vivo, the three mAbs stained B cells of several BALB/c mice clearly stronger than the isotype control; this differential staining was significantly reduced in FcgammaRIIB-deficient mice. The results indicate that the three mAbs recognize (a) planted antigen on viable cultured cells and (b) soluble autoantigen in vivo, leading to immune complexes that bind to FcgammaRIIB. Further experiments demonstrated that antinuc IgG2a could be eluted from splenocytes of a male BXSB lupus mouse. Hence, at least part of the extrinsic IgG2a(b) found on BXSB B cells may represent FcgammaRIIB-bound nuc-IgG2a(b) complexes.
Li
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Department of Immunology, School of Medicine, University of Tromsø, Tromsø, Norway.
Egorina
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Bertelsen
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Dahlen
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Hannestad
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D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003433:Crosses, Genetic; D005260:Female; D005434:Flow Cytometry; D007074:Immunoglobulin G / Q000276:immunology*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008814:Mice, Inbred NZB; D018345:Mice, Knockout; D009707:Nucleosomes / Q000276:immunology*; D017452:Receptors, IgG / Q000276:immunology*; D013154:Spleen / Q000276:immunology
D000974:Antibodies, Antinuclear; D000936:Antigen-Antibody Complex; C499172:Fcgr2b protein, mouse; D007074:Immunoglobulin G; D009707:Nucleosomes; D017452:Receptors, IgG