Volume 272, Number 20, Issue of May 16, 1997 pp. 12938-12944
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

High Affinity Saturable Uptake of Oxidized Low Density Lipoprotein by Macrophages from Mice Lacking the Scavenger Receptor Class A Type I/II

(Received for publication, November 5, 1996, and in revised form, February 3, 1997)

Marilee Lougheed , Corey Ming Lum , Wenhua Ling , Hiroshi Suzuki ^§ , Tatsuhiko Kodama ^§ and Urs Steinbrecher

From the  Department of Medicine, The University of British Columbia, Vancouver, British Columbia, V5Z 4E3 Canada and ^§ Department of Molecular Biology and Medicine, Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153, Japan

Oxidation of low density lipoproteins (LDL) has been implicated as a causal factor in the pathogenesis of atherosclerosis. Oxidized LDL has been found to exhibit numerous potentially atherogenic properties in vitro, including receptor-mediated uptake by macrophages. Oxidized LDL is a ligand for the class A scavenger receptor type I/II (SR-AI/II), but cross-competition studies with cultured macrophages suggested that there is an additional receptor(s) that is specific for oxidized LDL and that does not interact with acetyl LDL or other chemically modified LDL. A number of macrophage membrane proteins, including CD36, FcRII-B2, scavenger receptor BI, and macrosialin/CD68, have been found to bind to oxidized LDL in vitro and have been proposed as candidate oxidized LDL receptors. However, because of overlapping ligand specificity with the SR-AI/II, it has been difficult to evaluate the relative importance of these proteins in the uptake of oxidized LDL by macrophages. In the present report, we have studied the uptake and degradation of oxidized LDL by macrophages from mice in which the SR-AI/II gene had been disrupted. The uptake of acetyl LDL was reduced by more than 80% in macrophages from scavenger receptor knockout mice, confirming that most of the uptake of acetyl LDL by macrophages can be attributed to this receptor. In contrast, the uptake of extensively oxidized LDL was reduced by only 30% and showed high affinity, saturable uptake with apparent K_m of about 5 µg/ml, similar to that of the SR-AI/II. This indicates that about 70% of the uptake of oxidized LDL in macrophages is attributable to an alternate oxidized LDL receptor(s). In contrast to findings reported with CD36, mildly oxidized LDL was internalized much more slowly than extensively oxidized LDL. Unlabeled oxidized LDL, polyinosinic acid, phosphatidylserine-rich liposomes, and LDL or bovine albumin modified by fatty acid oxidation products were effective competitors for the uptake of radioiodinated oxidized LDL by macrophages from knockout mice, whereas acetyl LDL and malondialdehyde-modified LDL were relatively poor competitors. This ligand specificity differs from that of CD36-related (class B) scavenger receptors but is similar to the reported specificity of macrosialin/CD68 in ligand blots. However, the rate of uptake of oxidized LDL by knockout macrophages was not increased by phorbol ester or in thioglycollate-elicited macrophages, both of which are expected to increase the amount of macrosialin on the cell surface. In macrophages from SR-AI/II knockout mice, ligand blots of membrane proteins with iodinated, oxidized, or acetylated LDL revealed several bands, with apparent molecular size on SDS-polyacrylamide gel electrophoresis of 60, 94, 124, and 210 kDa, but none of the bands were specific for oxidized LDL. These results provide direct evidence that a receptor other than SR-AI/II is responsible for most of the uptake of oxidized LDL in murine macrophages, but further studies are needed to identify the receptor(s) involved.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S.-R. Ji, Y. Wu, L. Zhu, L. A. Potempa, F.-L. Sheng, W. Lu, and J. Zhao Cell membranes and liposomes dissociate C-reactive protein (CRP) to form a new, biologically active structural intermediate: mCRPm FASEB J, January 1, 2007; 21(1): 284 - 294. [Abstract] [Full Text] [PDF]

				A. M. Aslanian and I. F. Charo Targeted Disruption of the Scavenger Receptor and Chemokine CXCL16 Accelerates Atherosclerosis Circulation, August 8, 2006; 114(6): 583 - 590. [Abstract] [Full Text] [PDF]

				Y. Tamura, J.-i. Osuga, H. Adachi, R.-i. Tozawa, Y. Takanezawa, K. Ohashi, N. Yahagi, M. Sekiya, H. Okazaki, S. Tomita, et al. Scavenger Receptor Expressed by Endothelial Cells I (SREC-I) Mediates the Uptake of Acetylated Low Density Lipoproteins by Macrophages Stimulated with Lipopolysaccharide J. Biol. Chem., July 23, 2004; 279(30): 30938 - 30944. [Abstract] [Full Text] [PDF]

				H. Du, S. Schiavi, N. Wan, M. Levine, D. P. Witte, and G. A. Grabowski Reduction of Atherosclerotic Plaques by Lysosomal Acid Lipase Supplementation Arterioscler. Thromb. Vasc. Biol., January 1, 2004; 24(1): 147 - 154. [Abstract] [Full Text] [PDF]

				C.A. Argmann, C.G. Sawyez, C.J. McNeil, R.A. Hegele, and M.W. Huff Activation of Peroxisome Proliferator-Activated Receptor Gamma and Retinoid X Receptor Results in Net Depletion of Cellular Cholesteryl Esters in Macrophages Exposed to Oxidized Lipoproteins Arterioscler. Thromb. Vasc. Biol., March 1, 2003; 23(3): 475 - 482. [Abstract] [Full Text] [PDF]

				I. Suc, S. Brunet, G. Mitchell, G.-E. Rivard, and E. Levy Oxidative tyrosylation of high density lipoproteins impairs cholesterol efflux from mouse J774 macrophages: role of scavenger receptors, classes A and B J. Cell Sci., January 1, 2003; 116(1): 89 - 99. [Abstract] [Full Text] [PDF]

				V. V. Kunjathoor, M. Febbraio, E. A. Podrez, K. J. Moore, L. Andersson, S. Koehn, J. S. Rhee, R. Silverstein, H. F. Hoff, and M. W. Freeman Scavenger Receptors Class A-I/II and CD36 Are the Principal Receptors Responsible for the Uptake of Modified Low Density Lipoprotein Leading to Lipid Loading in Macrophages J. Biol. Chem., December 13, 2002; 277(51): 49982 - 49988. [Abstract] [Full Text] [PDF]

				C. A. Argmann, C. H. Van Den Diepstraten, C. G. Sawyez, J. Y. Edwards, R. A. Hegele, B. M. Wolfe, and M. W. Huff Transforming Growth Factor-{beta}1 Inhibits Macrophage Cholesteryl Ester Accumulation Induced by Native and Oxidized VLDL Remnants Arterioscler. Thromb. Vasc. Biol., December 1, 2001; 21(12): 2011 - 2018. [Abstract] [Full Text] [PDF]

				G. PUGLIESE, F. PRICCI, C. IACOBINI, G. LETO, L. AMADIO, P. BARSOTTI, L. FRIGERI, D. K. HSU, H. VLASSARA, F.-T. LIU, et al. Accelerated diabetic glomerulopathy in galectin-3/AGE receptor 3 knockout mice FASEB J, November 1, 2001; 15(13): 2471 - 2479. [Abstract] [Full Text] [PDF]

				V. R. Babaev, L. A. Gleaves, K. J. Carter, H. Suzuki, T. Kodama, S. Fazio, and M. F. Linton Reduced Atherosclerotic Lesions in Mice Deficient for Total or Macrophage-Specific Expression of Scavenger Receptor-A Arterioscler. Thromb. Vasc. Biol., December 1, 2000; 20(12): 2593 - 2599. [Abstract] [Full Text] [PDF]

				H. C McGill Jr, C A. McMahan, E. E Herderick, G. T Malcom, R. E Tracy, and J. P Strong Origin of atherosclerosis in childhood and adolescence Am. J. Clinical Nutrition, November 1, 2000; 72 (5): 1307S - 1315S. [Abstract] [Full Text] [PDF]

				E. A. Podrez, G. Hoppe, J. O'Neil, L. M. Sayre, N. Sheibani, and H. F. Hoff Macrophage receptors responsible for distinct recognition of low density lipoprotein containing pyrrole or pyridinium adducts: models of oxidized low density lipoprotein J. Lipid Res., September 1, 2000; 41(9): 1455 - 1463. [Abstract] [Full Text]

				V. Terpstra, E. S. van Amersfoort, A. G. van Velzen, J. Kuiper, and T. J. C. van Berkel Hepatic and Extrahepatic Scavenger Receptors : Function in Relation to Disease Arterioscler. Thromb. Vasc. Biol., August 1, 2000; 20(8): 1860 - 1872. [Full Text] [PDF]

				M. Butler, R. M. Crooke, M. J. Graham, K. M. Lemonidis, M. Lougheed, S. F. Murray, D. Witchell, U. Steinbrecher, and C. F. Bennett Phosphorothioate Oligodeoxynucleotides Distribute Similarly in Class A Scavenger Receptor Knockout and Wild-Type Mice J. Pharmacol. Exp. Ther., February 1, 2000; 292(2): 489 - 496. [Abstract] [Full Text]

				M. P. J. de Winther, K. W. van Dijk, L. M. Havekes, and M. H. Hofker Macrophage Scavenger Receptor Class A : A Multifunctional Receptor in Atherosclerosis Arterioscler. Thromb. Vasc. Biol., February 1, 2000; 20(2): 290 - 297. [Abstract] [Full Text] [PDF]

				M Kostich, A Fire, and D. Fambrough Identification and molecular-genetic characterization of a LAMP/CD68-like protein from Caenorhabditis elegans J. Cell Sci., January 7, 2000; 113(14): 2595 - 2606. [Abstract] [PDF]

				A. Gomez-Munoz, J. S. Martens, and U. P. Steinbrecher Stimulation of Phospholipase D Activity by Oxidized LDL in Mouse Peritoneal Macrophages Arterioscler. Thromb. Vasc. Biol., January 1, 2000; 20(1): 135 - 143. [Abstract] [Full Text] [PDF]

				H. Massaeli, J. A. Austria, and G. N. Pierce Chronic Exposure of Smooth Muscle Cells to Minimally Oxidized LDL Results in Depressed Inositol 1,4,5-Trisphosphate Receptor Density and Ca2+ Transients Circ. Res., September 17, 1999; 85(6): 515 - 523. [Abstract] [Full Text] [PDF]

				M. Lougheed, E. D. W. Moore, D. R. L. Scriven, and U. P. Steinbrecher Uptake of Oxidized LDL by Macrophages Differs From That of Acetyl LDL and Leads to Expansion of an Acidic Endolysosomal Compartment Arterioscler. Thromb. Vasc. Biol., August 1, 1999; 19(8): 1881 - 1890. [Abstract] [Full Text] [PDF]

				M. Febbraio, N. A. Abumrad, D. P. Hajjar, K. Sharma, W. Cheng, S. F. A. Pearce, and R. L. Silverstein A Null Mutation in Murine CD36 Reveals an Important Role in Fatty Acid and Lipoprotein Metabolism J. Biol. Chem., July 2, 1999; 274(27): 19055 - 19062. [Abstract] [Full Text] [PDF]

				S. C. Whitman, C. A. Argmann, C. G. Sawyez, D. B. Miller, R. A. Hegele, and M. W. Huff Uptake of type IV hypertriglyceridemic VLDL by cultured macrophages is enhanced by interferon-{gamma} J. Lipid Res., June 1, 1999; 40(6): 1017 - 1028. [Abstract] [Full Text]

				D. A. Bird, K. L. Gillotte, S. Horkko, P. Friedman, E. A. Dennis, J. L. Witztum, and D. Steinberg Receptors for oxidized low-density lipoprotein on elicited mouse peritoneal macrophages can recognize both the modified lipid moieties and the modified protein moieties: Implications with respect to macrophage recognition of apoptotic cells PNAS, May 25, 1999; 96(11): 6347 - 6352. [Abstract] [Full Text] [PDF]

				S. C. Whitman, S. L. Hazen, D. B. Miller, R. A. Hegele, J. W. Heinecke, and M. W. Huff Modification of Type III VLDL, Their Remnants, and VLDL From ApoE-Knockout Mice by p-Hydroxyphenylacetaldehyde, a Product of Myeloperoxidase Activity, Causes Marked Cholesteryl Ester Accumulation in Macrophages Arterioscler. Thromb. Vasc. Biol., May 1, 1999; 19(5): 1238 - 1249. [Abstract] [Full Text] [PDF]

				J. S. Martens, M. Lougheed, A. Gomez-Munoz, and U. P. Steinbrecher A Modification of Apolipoprotein B Accounts for Most of the Induction of Macrophage Growth by Oxidized Low Density Lipoprotein J. Biol. Chem., April 16, 1999; 274(16): 10903 - 10910. [Abstract] [Full Text] [PDF]

				T. Biwa, H. Hakamata, M. Sakai, A. Miyazaki, H. Suzuki, T. Kodama, M. Shichiri, and S. Horiuchi Induction of Murine Macrophage Growth by Oxidized Low Density Lipoprotein Is Mediated by Granulocyte Macrophage Colony-stimulating Factor J. Biol. Chem., October 23, 1998; 273(43): 28305 - 28313. [Abstract] [Full Text] [PDF]

				J. S. Martens, N. E. Reiner, P. Herrera-Velit, and U. P. Steinbrecher Phosphatidylinositol 3-Kinase Is Involved in the Induction of Macrophage Growth by Oxidized Low Density Lipoprotein J. Biol. Chem., February 27, 1998; 273(9): 4915 - 4920. [Abstract] [Full Text] [PDF]

				V. Terpstra, D. A. Bird, and D. Steinberg Evidence that the lipid moiety of oxidized low density lipoprotein plays a role in its interaction with macrophage receptors PNAS, February 17, 1998; 95(4): 1806 - 1811. [Abstract] [Full Text] [PDF]

				V. Terpstra, N. Kondratenko, and D. Steinberg Macrophages lacking scavenger receptor A show a decrease in binding and uptake of acetylated low-density lipoprotein and of apoptotic thymocytes, but not of oxidatively damaged red blood cells PNAS, July 22, 1997; 94(15): 8127 - 8131. [Abstract] [Full Text] [PDF]