Treatment with dsDNA-anti-dsDNA antibody complexes extends survival, decreases anti-dsDNA antibody production and reduces severity of nephritis in MRLlpr mice

Lupus nephritis results from the deposition on the glomerular basement membrane of antibodies cross-reacting with dsDNA. In an attempt to suppress the production of such antibodies in the MRLlpr mouse strain, mice were treated by injections of dsDNA-anti-dsDNA antibody complexes formed in an excess of syngeneic polyclonal antibodies to increase the immunogenicity of idiotypes. A first group of mice was treated after the onset of high-affinity anti-dsDNA IgG antibodies (3 months). We show here that fortnightly injections of dsDNA-anti-dsDNA antibody complexes significantly extend mouse survival over that of two control groups treated with either the carrier buffer or with sham complexes. Treated mice produced 5-fold less anti-dsDNA antibodies than control mice and presented a reduced nephritis activity index at the age of 7 months. Specific antiidiotypic antibody levels were not modified in the treated group, while showing a sharp decrease in the control group between months 6 and 7. Mice of a second group were started on injections of dsDNA-anti-dsDNA complexes at the age of 4 months when nephritis was already ongoing, and were followed until death while receiving fortnightly injections. Forty percent of the treated mice were still alive after one year, while none of the control mice survived. dsDNA-anti-dsDNA complexes have therefore the potential of reducing the production of anti-dsDNA antibody production and the severity of nephritis in MRLlpr mice. These findings could be relevant for the treatment of human lupus

Variable region heavy chain glycosylation determines the anticoagulant activity of a factor VIII antibody.

BACKGROUND: N-glycosylation occurs in the variable region of about 10% of antibodies but the role of carbohydrate at this location is still poorly understood. OBJECTIVES: We investigated the function of N-glycosylation in the variable region of the heavy chain of a human monoclonal antibody, mAb-LE2E9, that partially inhibits factor VIII (FVIII) activity during coagulation. METHODS AND RESULTS: Enzymatic deglycosylation indicated that the oligosaccharides do not determine the affinity of the antibody but enhance its FVIII neutralizing activity. A mutant antibody lacking the N-glycosylation site in the variable region of the heavy chain inhibited FVIII activity by up to 40%, while inhibition by the native antibody was 80%. To evaluate the physiological effect of such a FVIII inhibition, we investigated the ability of the mutant antibody devoid of N-glycosylation in the variable region to prevent thrombosis in mice with a strong prothombotic phenotype resulting from a type II deficiency mutation in the heparin binding site of antithrombin. Despite its moderate inhibition of FVIII activity, the mutant antibody significantly prevented thrombosis in treated animals. We also carried out glycan analysis of native and mutant antibodies. CONCLUSIONS: Modification of glycosylation in the variable region of antibodies contributes to the diversity of FVIII type II inhibition possibly by steric hindrance of the active site of FVIII by glycans, and may provide a novel strategy to modulate the functional activity of therapeutic antibodies