Correlation of antiphospholipid antibody recognition with the structure of synthetic oxidized phospholipids. Importance of Schiff base formation and aldol condensation.
The oxidation of low density lipoproteins (LDL) has been correlated with atherogenesis through a variety of pathways. The process involves nonspecific fragmentation, oxidative breakdown, and modification of the lipids and protein of LDL. The process yields a variety of bioactive products, including aldehyde-containing phospholipids, which can cross-react with primary amines (i.e. peptides or phospholipid head groups) to yield Schiff base products. We also demonstrate that such oxidized phospholipid products may further react through a post-oxidation chemical pathway involving aldol condensation. EO6, an IgM monoclonal autoantibody to oxidized phospholipids, blocks the uptake of oxidized LDL (OxLDL) by macrophages. Because the epitope(s) of EO6 also blocks the uptake of OxLDL, a series of oxidized phospholipids, their peptide complexes, and their aldol condensates have been synthesized and characterized, and their antigenicity has been determined. This study defines structural motifs of oxidized phospholipids responsible for antigenicity for EO6. Certain monomeric phospholipids containing short chain fatty acids were antigenic whether oxidized or not in the sn-2 position. However, oxidized phospholipids containing sn-1 long chain fatty acids were not antigenic unless the sn-2 oxidized fatty acid contained an aldehyde that first reacted with a peptide yielding a Schiff base or the sn-2 oxidized fatty acid underwent an aldol type self-condensation. Our data indicate that the phosphorylcholine head group is essential for antigenicity, but its availability depends on the oxidized phospholipid conformation. We suggest that upon oxidation, similar reactions occur in phospholipids on the surface of LDL, generating ligands for macrophage recognition. Synthetic imine adducts of oxidized phospholipids of this type are capable of blocking the uptake of OxLDL.
Friedman
Peter
P
Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093-0601, USA.
Horkko
Sohvi
S
Steinberg
Daniel
D
Witztum
Joseph L
JL
Dennis
Edward A
EA
D017152:Antibodies, Antiphospholipid / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004305:Dose-Response Relationship, Drug; D005227:Fatty Acids / Q000737:chemistry; D006433:Hemocyanins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007075:Immunoglobulin M / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008055:Lipids / Q000737:chemistry; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D008264:Macrophages / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D010100:Oxygen / Q000378:metabolism*; D010742:Phospholipid Ethers / Q000737:chemistry; D010743:Phospholipids / Q000378:metabolism*; D010767:Phosphorylcholine / Q000737:chemistry
C400184:1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine; D017152:Antibodies, Antiphospholipid; D000911:Antibodies, Monoclonal; D005227:Fatty Acids; D007075:Immunoglobulin M; D008055:Lipids; D008077:Lipoproteins, LDL; D010742:Phospholipid Ethers; D010743:Phospholipids; D010767:Phosphorylcholine; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D008239:Lysine; D010100:Oxygen