title: Histone-containing immune complexes are to a large extent responsible for anti-dsDNA reactivity in the Farr assay of active SLE patients.
abstract: Increased titres of anti-dsDNA antibodies, especially if of high avidity, are associated with renal exacerbations in patients with systemic lupus erythematosus (SLE). One of the most reliable assays to measure anti-dsDNA antibodies, the Farr assay, is believed to detect preferentially high avidity antibodies. Purified non-complexed monoclonal antibodies (mAbs) against nucleosomes, obtained from mice with SLE, are not reactive in the Farr assay, but can become so once complexed to nucleosomes. These Farr-positive, nucleosome containing, immune complexes were also able to bind in vivo to the glomerular basement membrane (GBM), predominantly via heparan sulphate (HS). To evaluate whether in SLE patients the same kind of immune complexes are responsible for Farr reactivity, IgG from serum or plasma was isolated under dissociating and physiological conditions. We observed that after purification under dissociating conditions, Farr reactivity was significantly decreased (P<0.0001) in contrast to reactivity with histones and two 'control' antigens: Epstein Barr Virus (EBV) and Ro/SS-A. Reactivity with nucleosomes also decreased after purification, although to a lesser extent. Plasma purified under physiological conditions showed no decrease in Farr reactivity. The importance of histones for the generation of immune complexes is supported by the two following observations. Firstly, the presence of histones could be demonstrated in serum and plasma of SLE patients but not in serum of healthy controls or in IgG preparations purified under dissociating conditions. Secondly, Farr reactivity of purified IgG preparations could be restored by addition of purified histones. From these studies we conclude that histones containing immune complexes are responsible for a large part of the Farr reactivity in active SLE, and are therefore indirectly implicated in the pathogenesis of lupus nephritis.
authors.affiliation: Department of Autoimmune Diseases, CLB, Amsterdam, The Netherlands.
authors.forename: M N
authors.initials: MN
authors.lastname: Hylkema
authors.forename: M C
authors.initials: MC
authors.lastname: van Bruggen
authors.forename: T
authors.initials: T
authors.lastname: ten Hove
authors.forename: J
authors.initials: J
authors.lastname: de Jong
authors.forename: A J
authors.initials: AJ
authors.lastname: Swaak
authors.forename: J H
authors.initials: JH
authors.lastname: Berden
authors.forename: R J
authors.initials: RJ
authors.lastname: Smeenk
mesh_terms: D000818:Animals; D000974:Antibodies, Antinuclear / Q000032:analysis / Q000097:blood*; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000097:blood*; D001324:Autoantigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004854:Herpesvirus 4, Human / Q000276:immunology; D006657:Histones / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood; D007075:Immunoglobulin M / Q000032:analysis / Q000097:blood; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D051379:Mice; D020733:RNA, Small Cytoplasmic; D015531:Radioimmunoprecipitation Assay; D012261:Ribonucleoproteins / Q000276:immunology; D000099207:SS-A Antigen
chemical_list: D000974:Antibodies, Antinuclear; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D001324:Autoantigens; D006657:Histones; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human