J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.M600158-JLR200 on October 5, 2006

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Journal of Lipid Research, Vol. 48, 19-29, January 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Novel anti-cholesterol monoclonal immunoglobulin G antibodies as probes and potential modulators of membrane raft-dependent immune functions

Adrienn Bíró1,*,{dagger}, László Cervenak1,{dagger}, Andrea Balogh1,§, András Lorincz§, Katalin Uray**, Anna Horváth*, László Romics*,{dagger}, János Matkó§, George Füst*,{dagger} and Glória László2,§

* Third Department of Internal Medicine, Research Laboratory, Semmelweis University, Budapest, Hungary
{dagger} Research Group on Metabolism and Atherosclerosis, Hungarian Academy of Sciences-Semmelweis University, Budapest, Hungary
§ Department of Immunology, Eötvös Lorand University, Budapest, Hungary
** Department of Organic Chemistry, Eötvös Lorand University, Budapest, Hungary

Published, JLR Papers in Press, October 5, 2006.

1 A. Bíró, L. Cervenak, and A. Balogh contributed equally to this work.

2 To whom correspondence should be addressed. e-mail: glorial{at}elte.hu

Natural autoantibodies against cholesterol are present in the sera of all healthy individuals; their function, production, and regulation, however, are still unclear. Here, we managed to produce two monoclonal anti-cholesterol antibodies (ACHAs) by immunizing mice with cholesterol-rich liposomes. The new ACHAs were specific to cholesterol and to some structurally closely related 3ß-hydroxyl sterols, and they reacted with human lipoproteins VLDL, LDL, and HDL. They bound, usually with low avidity, to live human or murine lymphocyte and monocyte-macrophage cell lines, which was enhanced substantially by a moderate papain digestion of the cell surface, removing some protruding extracellular protein domains. Cell-bound ACHAs strongly colocalized with markers of cholesterol-rich lipid rafts and caveolae at the cell surface and intracellularly with markers of the endoplasmic reticulum and Golgi complex. These data suggest that these IgG ACHAs may serve as probes of clustered cholesterol (e.g., different lipid rafts) in live cells and thus may also have immunomodulatory potential.

Supplementary key words membrane cholesterol • lymphocytes • monocyte-macrophage cells • lipoproteins • lipid rafts • caveolae


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