title: Foam cell formation containing lipid droplets enriched with free cholesterol by hyperlipidemic serum.
abstract: A monoclonal antibody, ASH1a/256C (256C), which binds to atherosclerotic lesions in Watanabe heritable hyperlipidemic rabbit (WHHL) aorta in vivo, recognizes complex structures of phosphatidylcholine mixed with neutral lipids. In the present study, a cell culture system is described in which foam cells express 256C-positive lipid droplets. J774.1 macrophages were incubated in the presence of a small volume of WHHL serum for 24 h to produce foam cells, which were then incubated without the WHHL serum for 3 days. Oil red O-positive lipid droplets appeared on day 1, and were present in the cells during the whole incubation period. The lipid droplets in the cells were positively immunostained with antibody 256C on day 4, although they were negative on day 1. Expression of the antigenic lipid droplets was also induced by the addition of acetylated LDL or sera from patients with hyperlipidemia. When foam cells were induced by the addition of WHHL serum, cellular content of cholesteryl ester was greatly increased but then decreased to near basal levels by day 4. Concomitantly, cellular free cholesterol increased during the culture period, indicating that the cholesteryl ester changes to free cholesterol by day 4. The lipid droplets in the foam cells on day 4 were positively stained with filipin, a fluorescent probe for free cholesterol, as well as with 256C antibody, indicating that free cholesterol is enriched in antigenic lipid droplets. These observations suggest that hydrolysis and rearrangement of cellular cholesterol take place in foam cells to form complex structures of phosphatidylcholine and free cholesterol in lipid droplets.
authors.affiliation: Department of Molecular Pathology, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Tsukui, Kanagawa 199-0195, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Itabe
authors.forename: Y
authors.initials: Y
authors.lastname: Higashi
authors.forename: Y
authors.initials: Y
authors.lastname: Fujimoto
authors.forename: M
authors.initials: M
authors.lastname: Shiomi
authors.forename: M
authors.initials: M
authors.lastname: Yoshizumi
authors.forename: Y
authors.initials: Y
authors.lastname: Ouchi
authors.forename: T
authors.initials: T
authors.lastname: Takano
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002784:Cholesterol / Q000032:analysis* / Q000378:metabolism; D002788:Cholesterol Esters / Q000032:analysis / Q000378:metabolism; D003300:Copper / Q000494:pharmacology; D005372:Filipin; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D005487:Foam Cells / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology*; D006949:Hyperlipidemias / Q000097:blood*; D007700:Kinetics; D050356:Lipid Metabolism; D015227:Lipid Peroxidation; D008055:Lipids / Q000032:analysis*; D008077:Lipoproteins, LDL / Q000378:metabolism / Q000494:pharmacology; D008264:Macrophages; D051379:Mice; D010713:Phosphatidylcholines / Q000378:metabolism; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D002788:Cholesterol Esters; D005456:Fluorescent Dyes; D008055:Lipids; D008077:Lipoproteins, LDL; D010713:Phosphatidylcholines; C032629:acetyl-LDL; C099252:oxidized low density lipoprotein; D003300:Copper; D005372:Filipin; D002784:Cholesterol