title: Association of autoantibodies to nuclear lamin B1 with thromboprotection in systemic lupus erythematosus: lack of evidence for a direct role of lamin B1 in apoptotic blebs.
abstract: OBJECTIVE
  To demonstrate the association between autoantibodies to nuclear lamin B1 (aLB1) and protection against thrombosis ("thromboprotection") in patients with systemic lupus erythematosus (SLE), and to elucidate the mechanism by which aLB1 cause thromboprotection in vivo. Since a number of autoantigens in SLE have been localized specifically to the external surface of apoptotic blebs, it was hypothesized that circulating aLB1 may block the procoagulant effect of apoptotic blebs by binding to LB1 displayed at the external bleb surface.
  
  
  METHODS
  A cross-sectional study was performed using serum samples obtained at first evaluation of 259 English Canadian and French Canadian patients from SLE registries at 3 hospitals. A case-control study was performed to analyze the relationship between aLB1 and lupus anticoagulant (LAC) status and thrombotic manifestations between onset of disease and last followup. Reactivity of aLB1 with Jurkat or endothelial cells, which had been induced to undergo apoptosis, was determined by indirect immunofluorescence. Localization of LB1 in apoptotic cells and blebs was analyzed by confocal microscopy and surface labeling of cell membrane proteins.
  
  
  RESULTS
  High-titer aLB1 was restricted to a subset of SLE patients (46 patients), with an overall frequency of 17.8% (range 11.6-24.3% in the 3 centers). LB1 antibodies were significantly associated with LAC but not with antibodies to cardiolipin (aCL) or beta(2)-glycoprotein I (anti-beta(2)GPI). The frequency of thrombosis differed markedly depending on aLB1 and LAC status, as follows: presence of LAC and absence of aLB1 50%, presence of both LAC and aLB1 22.7%, absence of both LAC and aLB1 25.5%, absence of LAC and presence of aLB1, 20.8%. Further subclassification of patients based on aCL and anti-beta(2)GPI status revealed that, in the presence of LAC but in the absence of aCL, anti-beta(2)GPI, and aLB1, the frequency of thrombosis was 40%, whereas in the presence of aLB1, it decreased strikingly, to 9.1%. LB1 was found to be translocated into surface membrane blebs during apoptosis and to be entirely enclosed within the apoptotic bleb plasma membrane of Jurkat and endothelial cells.
  
  
  CONCLUSIONS
  The presence of aLB1 in SLE patients with LAC essentially nullifies the strong prothrombotic risk associated with LAC. Hence, aLB1 is associated with thromboprotection. Reactivity of aLB1 with apoptotic blebs does not seem to play a direct role in mediating this protection, since LB1 is buried within apoptotic blebs and inaccessible to circulating aLB1. The mechanism by which aLB1 confers thromboprotection in SLE remains to be elucidated.
authors.affiliation: Autoimmunity Research Laboratory, Notre-Dame Hospital, Centre Hospitalier de l'Université de Montréal, 1560 Sherbrooke Street East, Montreal, Quebec H2L 4M1, Canada.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Dieudé
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Senécal
authors.forename: Joyce
authors.initials: J
authors.lastname: Rauch
authors.forename: John G
authors.initials: JG
authors.lastname: Hanly
authors.forename: Paul
authors.initials: P
authors.lastname: Fortin
authors.forename: Nathalie
authors.initials: N
authors.lastname: Brassard
authors.forename: Yves
authors.initials: Y
authors.lastname: Raymond
mesh_terms: D000328:Adult; D000368:Aged; D001324:Autoantigens / Q000276:immunology / Q000378:metabolism; D016022:Case-Control Studies; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003430:Cross-Sectional Studies; D003593:Cytoplasm / Q000378:metabolism; D053938:DNA Fragmentation / Q000276:immunology; D005260:Female; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D019169:Jurkat Cells / Q000276:immunology / Q000378:metabolism; D034921:Lamin Type B / Q000276:immunology* / Q000378:metabolism; D016682:Lupus Coagulation Inhibitor / Q000097:blood*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D013927:Thrombosis / Q000276:immunology*; D053482:beta 2-Glycoprotein I
chemical_list: D001324:Autoantigens; D006023:Glycoproteins; D007074:Immunoglobulin G; D034921:Lamin Type B; D016682:Lupus Coagulation Inhibitor; D053482:beta 2-Glycoprotein I