Heparin and heparinoids prevent the binding of immune complexes containing nucleosomal antigens to the GBM and delay nephritis in MRL/lpr mice.
Monoclonal anti-nucleosome antibodies (mAbs) complexed to nucleosomal antigens can bind to DNA and to heparan sulfate (HS) in ELISA and to the GBM in vivo in a rat renal perfusion system, whereas non-complexed mAbs do not bind [1]. In this study, we analyzed whether heparin (HEP) or N-desulfated/acetylated heparins (DSA-HEP), structurally and functionally strongly related to HS, are able to prevent the binding of these complexed mAbs to DNA and to HS in vitro and to rat GBM in vivo. In ELISA the binding of nucleosome complexed antinucleosome antibodies to DNA and HS was inhibited dose-dependently by HEP, DSA-HEP and low molecular weight (LMW) DSA-HEP. Intravenous injection of nucleosome/anti-nucleosome immune complexes without heparin/heparinoids in BALB/c mice led to GBM binding, while simultaneous injection of heparin/heparinoids with complexed antibodies or pretreatment with heparin subcutaneously prior to injection of complexes prevented this binding. Subsequently, we tested the preventive effect of HEP, DSA-HEP and LMW-DSA-HEP on progression of renal disease in MRL/lpr mice. Treatment was started at an age of eight weeks in a dose of 50 micrograms daily. With all three drugs albuminuria was significantly delayed compared to PBS treated controls (cumulative incidence of proteinuria at 20 weeks in controls 60% vs. 13%, 14% and 6% respectively for HEP, DSA-HEP and LMW-DSA-HEP; P < 0.05). At week 21 the glomerulonephritis was histologically less severe in heparin/heparinoid treated animals (P = 0.02). In immunofluorescence the amount of immunoglobulin and C3 deposits in the glomerular capillary wall tended to be less in heparin/heparinoid treated mice compared to PBS treated controls (P = 0.07). Furthermore, at 20 weeks anti-HS levels in plasma of heparin/heparinoid treated mice were significantly lower (P < 0.05). We conclude that interaction of heparin or heparin analogs with HS reactive immune complexes containing nucleosomal antigens prevents the binding of these immune complexes to the GBM and delays nephritis in MRL/lpr mice.
Department of Pathology, University Hospital Nijmegen, The Netherlands.
M C
MC
van Bruggen
B
B
Walgreen
T P
TP
Rijke
M J
MJ
Corsius
K J
KJ
Assmann
R J
RJ
Smeenk
G W
GW
van Dedem
K
K
Kramers
J H
JH
Berden
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D000936:Antigen-Antibody Complex / Q000187:drug effects / Q000276:immunology*; D000937:Antigen-Antibody Reactions / Q000187:drug effects; D001323:Autoantibodies / Q000276:immunology; D001485:Basement Membrane / Q000187:drug effects / Q000276:immunology; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006493:Heparin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology; D007678:Kidney Glomerulus / Q000187:drug effects / Q000276:immunology*; D008181:Lupus Nephritis / Q000276:immunology / Q000517:prevention & control*; D051379:Mice; D019463:Mice, Inbred MRL lpr; D009707:Nucleosomes / Q000187:drug effects / Q000276:immunology*
D000925:Anticoagulants; D000936:Antigen-Antibody Complex; D001323:Autoantibodies; D006495:Heparin, Low-Molecular-Weight; D009707:Nucleosomes; D006493:Heparin; D004247:DNA