Abstract 15: Serum Amyloid A is Not Just an HDL-Associated Lipoprotein

Abstract

Serum Amyloid A (SAA) is traditionally thought to be only found with HDL; however, recently several groups have found that SAA is found on apoB-containing lipoproteins (LDL and VLDL) under some circumstances. The goal of this study was to determine the relative lipoprotein association of SAA. Native human acute phase SAA was isolated from acute phase plasma collected from patients undergoing cardiovascular bypass surgery; the SAA was purified and delipidated. Plasma was collected from a group of healthy, non-obese humans with low levels of SAA (< 2 mg/L) and lipoproteins (VLDL, LDL and HDL) were isolated by density gradient ultracentrifugation. The delipidated SAA (at 2 concentrations) was incubated in vitro with the various lipoprotein preparations then samples were re-isolated by fast protein liquid chromatography and SAA was analyzed by ELISA and immunoblot. When SAA was incubated with any single lipoprotein all of the SAA was found associated with that lipoprotein and none remained in a lipid-free form. When SAA was incubated with a mixture of VLDL, LDL and HDL (based on equal protein and corresponding to concentrations in plasma) a majority of SAA (50-60%) was found on HDL with the remaining SAA found on VLDL and LDL. When SAA first complexed to HDL was added to a mixture of SAA-free LDL and VLDL the majority of SAA remained with the HDL (76-86%) but 5-10% of the SAA was found on each of LDL and VLDL. When SAA first complexed to either apoB-containing lipoprotein was then added to a mixture of HDL and the other apoB-containing lipoprotein most SAA moved to HDL (55-70%) but the remainder was found on the apoB-containing particles. Thus, SAA can move between lipoprotein particles in vitro. To determine if the presence of SAA on apoB-containing lipoproteins had a functional effect we evaluated lipoprotein-proteoglycan binding affinity. Proteoglycan mediated lipoprotein retention in the vessel wall is thought to be one of the key steps in initiation of atherosclerosis. Compared to SAA-free LDL or VLDL, the presence of SAA on apoB-containing lipoproteins caused increased proteoglycan binding affinity. Thus, SAA is not simply an HDL lipoprotein, but SAA can move between lipoprotein particles, and the presence of SAA on apoB particles may increase their atherogenicity.