J. Lipid Res.
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Journal of Lipid Research, Vol. 42, 1771-1781, November 2001
Copyright © 2001 by Lipid Research, Inc.

Foam cell formation containing lipid droplets enriched with free cholesterol by hyperlipidemic serum

Masahiro Moria, Hiroyuki Itabea, Yusuke Higashia, Yasuyuki Fujimotoa, Masahiro Shiomib, Masao Yoshizumic, Yasuyoshi Ouchic, and Tatsuya Takanoa
a Department of Molecular Pathology, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Tsukui, Kanagawa 199-0195, Japan
b Institute of Experimental Animals, Kobe University School of Medicine, Kobe, Japan
c Department of Geriatric Medicine, Graduate School of Medicine, Tokyo University School of Medicine, Tokyo, Japan

Correspondence to: Tatsuya Takano, To whom correspondence should be addressed., t_takano{at}pharm.teikyo-u.ac.jp (E-mail)

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. — Mori, M., H. Itabe, Y. Higashi, Y. Fujimoto, M. Shiomi, M. Yoshizumi, Y. Ouchi, and T. Takano. Foam cell formation containing lipid droplets enriched with free cholesterol by hyperlipidemic serum. J. Lipid Res. 2001. 42: 1771–1781.

Supplementary key words: atherosclerosis, lipoproteins, macrophages, monoclonal antibody, phosphatidylcholine, WHHL rabbit


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