Anticardiolipin antibodies from patients with the antiphospholipid antibody syndrome recognize epitopes in both beta(2)-glycoprotein 1 and oxidized low-density lipoprotein.
BACKGROUND
We recently suggested that many anticardiolipin antibodies bind only to oxidized cardiolipin (OxCL) and/or to OxCL-beta(2)-glycoprotein 1 (beta(2)GP1) adducts but not to a "reduced" cardiolipin that is unable to undergo oxidation. To test this hypothesis, we investigated 24 sera, 4 protein A-purified IgG fractions, and 3 human monoclonal antibodies that were all isolated from patients with antiphospholipid antibody syndrome (APS); testing was also performed in 7 controls. Two monoclonal antibodies (IS3 and IS4) were selected for binding to CL and one was selected for binding to beta(2)GP1 (LJB8).


RESULTS
By chemiluminescent immunoassay, all APS sera samples bound only to OxCL and not to reduced CL, and the binding was inhibited >95% by OxCL but not reduced CL. All purified IgG fractions bound to beta(2)GP1 but only when the beta(2)GP1 was plated on microtiter wells coated with OxCL. All 3 monoclonal antibodies bound only to OxCL. On Western blots, IS4 and LJB8 bound to beta(2)GP1 as well as to delipidated apoB of oxidized LDL but not to native apoB. IS3 also bound to oxidized apoB on Western blot. Covalent modification of beta(2)GP1 with oxidation products of CL made it more antigenic for APS serum samples, for purified IgG fractions, and for the monoclonal antibodies.


CONCLUSIONS
These data support the hypothesis that oxidation of CL is needed to generate epitopes for many anticardiolipin antibodies and that some of these epitopes are covalent adducts of OxCL with beta(2)GP1 or apoB.
Hörkkö
S
S
Department of Medicine, University of California, San Diego, USA. shorkko@ucsd.edu
Olee
T
T
Mo
L
L
Branch
D W
DW
Woods
V L
VL
Palinski
W
W
Chen
P P
PP
Witztum
J L
JL
D017153:Antibodies, Anticardiolipin / Q000097:blood*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology*; D016736:Antiphospholipid Syndrome / Q000097:blood / Q000276:immunology*; D001055:Apolipoproteins B / Q000378:metabolism; D001667:Binding, Competitive / Q000276:immunology; D002308:Cardiolipins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007118:Immunoassay; D008077:Lipoproteins, LDL / Q000276:immunology* / Q000378:metabolism; D008163:Luminescent Measurements; D046911:Macromolecular Substances; D008297:Male; D010084:Oxidation-Reduction; D053482:beta 2-Glycoprotein I
D017153:Antibodies, Anticardiolipin; D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D002308:Cardiolipins; D000939:Epitopes; D006023:Glycoproteins; D008077:Lipoproteins, LDL; D046911:Macromolecular Substances; D053482:beta 2-Glycoprotein I; C099252:oxidized low density lipoprotein