title: Model IgG monoclonal autoantibody-anti-idiotype pair for dissecting the humoral immune response to oxidized low density lipoprotein.
abstract: Increasing evidence implicates IgG autoantibodies against oxidized forms of low density lipoprotein (oxLDL) in the pathophysiology of atherosclerotic arterial disease. However, insufficient knowledge of their structure and function is a key gap. Using an elderly LDL receptor-deficient atherosclerotic mouse, we isolated a novel IgG3k against oxLDL (designated MAb LO1). LO1 reacts with copper-oxidized LDL, but minimally with native LDL. Further analysis showed that MAb LO1 also reacts in vitro with malondialdehyde-conjugated LDL (MDA-LDL), a known key epitope in copper-oxidized LDL preparations. By screening a phage library expressing single chain variable region antibodies (scFv), we selected an anti-idiotype scFv (designated H3) that neutralizes MAb LO1 binding to MDA-LDL. Amino acid substitutions between H3 and an irrelevant control scFv C12 showed that residues in the H3 CDRH2, CDRH3, and CDRL2 are all critical for MAb LO1 binding, consistent with a conformational epitope on H3 involving both heavy and light chains. Comparison of amino acids in H3 CDRH2 and CDRL2 with apoB, the major LDL protein, showed homologous sequences, suggesting H3 has structural similarities to the MAb LO1 binding site on MDA-LDL. Immunocytochemical staining showed that MAb LO1 binds epitopes in mouse and human atherosclerotic lesions. The MAb LO1-H3 combination therefore provides a very promising model for analyzing the structure and function of an individual IgG autoantibody in relation to atherosclerosis.
authors.affiliation: Vascular Sciences Section, Department of Medicine, Imperial College, Hammersmith Hospital, London, United Kingdom.
authors.forename: Shang-Hung
authors.initials: SH
authors.lastname: Chang
authors.forename: Michael
authors.initials: M
authors.lastname: Johns
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Boyle
authors.forename: Ellen
authors.initials: E
authors.lastname: McConnell
authors.forename: Paul A
authors.initials: PA
authors.lastname: Kirkham
authors.forename: Colin
authors.initials: C
authors.lastname: Bicknell
authors.forename: M
authors.initials: M
authors.lastname: Zahoor-ul-Hassan Dogar
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Edwards
authors.forename: Oliver
authors.initials: O
authors.lastname: Gale-Grant
authors.forename: Ramzi
authors.initials: R
authors.lastname: Khamis
authors.forename: Kurrun V V
authors.initials: KV
authors.lastname: Ramkhelawon
authors.forename: Dorian O
authors.initials: DO
authors.lastname: Haskard
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D050197:Atherosclerosis / Q000276:immunology / Q000473:pathology; D001323:Autoantibodies / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D056724:Immunity, Humoral; D007074:Immunoglobulin G / Q000276:immunology*; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D008077:Lipoproteins, LDL / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D057127:Single-Chain Antibodies / Q000276:immunology*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007074:Immunoglobulin G; D007130:Immunoglobulin Idiotypes; D008077:Lipoproteins, LDL; D057127:Single-Chain Antibodies; C099252:oxidized low density lipoprotein