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Preparation and biodistribution of 99mtechnetium labelled oxidized LDL in man

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      Abstract

      Radiolabelled autologous low density lipoprotein (LDL) has previously been used to study in vivo distribution and metabolism of native-LDL. Non-invasive imaging of atherosclerotic lesions using 99mTc-LDL was shown to be feasible in animal models and patients but the clinical utility remains to be assessed. Since recent reports suggest that oxidized LDL may play a major role in the pathogenesis of atherosclerosis, we developed a technique to oxidize autologous LDL and compared the biodistribution of oxidized-LDL with that of native-LDL in man. In addition, we evaluated the uptake in vivo of oxidized- and native-LDL by atherosclerotic plaques. LDL, obtained from human plasma was treated with various combinations of copper ions and H2O2 to induce oxidative modification by increasing the content of lipid peroxidation products and electrophoretic mobility. When LDL (0.3 mg/ml) was incubated with 100 μM Cu2+ and 500 μM H2O2 oxidation occurred rapidly within 1 h, and was labelled with 99mTc efficiently as native LDL. In vivo distribution studies revealed a faster plasma clearance of oxidized-LDL compared to native-LDL, and a higher uptake by the reticuloendothelial system. Tomographic scintigraphy of the neck in patients suffering from transient ischemic attacks, revealed accumulation of radiolabelled LDL preparations in the carotid artery affected by atherosclerotic lesions. We developed a technique to rapidly oxidize LDL using copper and H2O2. Biodistribution data demonstrate that oxidized-LDL is rapidly cleared from circulation, is taken up mostly by organs rich in macrophages, and can be detected at the level of carotid plaques.

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      References

        • Lees RS
        • Garabedian HD
        • Lees AM
        • Schumacher DJ
        • Miller A
        • Isaacsohn JL
        • Derksen A
        • Strauss HW
        Technetium-99m low density lipoproteins: preparation and biodistribution.
        J Nucl Med. 1985; 26: 1056
        • Vallabhajosula S
        • Paidi M
        • Badimon JJ
        • Le N-A
        • Goldsmith SJ
        • Fuster V
        • Ginsberg HN
        Radiotracers for low density lipoprotein biodistribution studies in vivo: technetium-99m low density lipoprotein versus radioiodinated low density lipoprotein preparation.
        J Nucl Med. 1988; 29: 1237
        • Rosen MJ
        • Butler SP
        • Meinken GE
        • Wang TTS
        • Ramakrishnan R
        • Srivastava SC
        • Alderson PO
        • Ginsberg HN
        Indium-111-labeled LDL: a potential agent for imaging atherosclerotic disease and lipoprotein biodistribution.
        J Nucl Med. 1990; 31: 343
        • Lees AM
        • Lees RS
        • Schoen FJ
        • Isaacsohn JL
        • Fischman AJ
        • McKusic KA
        • Strauss HW
        Imaging human atherosclerosis with 99mTc-labeled low density lipoproteins.
        Arteriosclerosis. 1988; 8: 461
        • Lees RS
        • Lees AM
        • Strauss HW
        External imaging of human atherosclerosis.
        J Nucl Med. 1983; 24: 154
        • Vallabhajosula S
        • Goldsmith SJ
        99mTc-low density lipoprotein: intracellularly trapped radiotracer for non-invasive imaging of low density lipoprotein metabolism in vivo.
        in: 7th edn. Semin Nucl Med. XX. 1990: 68
        • Shih I-L
        • Lees RS
        • Chang MY
        • Lees AM
        Focal accumulation of an apolipoprotein B-based synthetic oligopeptide in the healing rabbit arterial wall.
        in: 7th edn. Proc Natl Acad Sci USA. 87. 1990: 1436
        • Fowler S
        • Shio H
        • Haley WJ
        Characterization of lipidladen aortic cells from cholesterol-fed rabbits. IV. Investigation of macrophage-like properties of aortic cell populations.
        Lab Invest. 1979; 4: 372
        • Schaffner T
        • Taylor K
        • Bartucci EJ
        • Fischer-Dzoga K
        • Benson JH
        • Glagov S
        • Wissler R
        Arterial foam cells with distinctive immunomorphologic and istochemical features of macrophages.
        Am J Pathol. 1980; 100: 57
        • Gerrity RG
        The role of the monocyte in atherogenesis. I. Transition of blood-borne monocytes into foam cells in fatty lesions.
        Am J Pathol. 1981; 103: 181
        • Gerrity RG
        The role of the monocyte in atherogenesis. II. Migration of foam cells from atherosclerotic lesions.
        Am J Pathol. 1981; 103: 191
        • Faggiotto A
        • Ross R
        • Harker L
        Studies of hypercholesterolemia in the non-human primate. I. Changes that lead to fatty streak formation.
        Arteriosclerosis. 1984; 4: 323
        • Rosenfeld ME
        • Tsukada T
        • Gown AM
        • Ross R
        Fatty streak initiation in Watanabe Heritable Hyperlipemic and comparably hypercholesterolemic fat-fed rabbits.
        Arteriosclerosis. 1987; 1: 9
        • Goldstein JL
        • Ho JK
        • Basu SK
        • Brown MS
        Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition.
        in: 7th edn. Proc Natl Acad Sci USA. 76. 1979: 333
        • Steinberg D
        • Parthasarathy S
        • Carew TE
        • Khoo JC
        • Witzum JL
        Beyond cholesterol. Modifications of low density lipoproteins that increases its atherogenicity.
        New Engl J Med. 1989; 320: 915
        • Steinbrecher UP
        • Lougheeed M
        • Kwan W-C
        • Dirks M
        Recognition of oxidized low density lipoprotein by the scavenger receptor of macrophages results from derivatization of apolipoprotein B by products of fatty acid peroxidation.
        J Biol Chem. 1989; 264: 15216
        • Hatch FT
        • Lees RS
        Practical methods for plasma lipoprotein analysis.
        Adv Lipid Res. 1968; 6: 1
        • Smith PK
        • Krohn RI
        • Hermanson GT
        • Mallia AK
        • Gartner FH
        • Provenzano MD
        • Fujimoto EK
        • Goeke NM
        • Olson BJ
        • Klent DC
        Measurement of protein using bicinchoninic acid.
        Anal Biochem. 1985; 150: 76
        • Iuliano L
        • Praticò D
        • Ghiselli A
        • Bonavita MS
        • Violi F
        Reaction of dipyrimadole with the hydroxyl radical.
        Lipids. 1992; 27: 349
        • Bull AW
        • Marnett LJ
        Determination of malondialdehyde by ion-pairing high performance liquid chromatography.
        Anal Biochem. 1985; 149: 284
        • Habeeb AFSA
        Determination of free amino groups in proteins by trinitrobenzenesulfonic acid.
        Anal Biochem. 1966; 14: 328
        • Esterbauer H
        • Gebicki J
        • Puhl H
        • Jurgens G
        The role of lipid peroxidation and antioxidants in oxidative modification of LDL.
        Free Rad Biol Med. 1992; 13: 341
        • Prat L
        • Carrio I
        • Roca M
        • Blasi J
        • Riembau V
        • Berna c
        • Torres G
        • Duncker D
        • Estorch M
        111In-polyclonal lg G and 125I-LDL accumulation in experimental arterial wall injury.
        in: Martin-Comin J Thakur ML Berna C Roca M Lomena F Radiolabelled Blood Elements. Recent Advances in Techniques and Applications. Plenum Press, New York, NY1994: 173
        • Virgolini I
        • Mülle C
        • Fitscha P
        • Chiba P
        • Sinzinger H
        Radiolabelling autologous monocytes with 111In-oxine for reinjection in patients with atherosclerosis.
        Prog Clin Biol Res. 1990; 355: 271
        • Praza L
        • Roca M
        • Rubio J
        • Ferre X
        • Richard Y
        • Mairal L
        • Martin-Comin J
        111In-platelet and 99mTc-human polyclonal immunoglobulin (HIG) scintigraphy in patients with cerebrovascular disease.
        in: Martin-Comin J Thakur ML Berna C Roca M Lomena F Radiolabelled Blood Elements. Recent Advances in Techniques and Applications. Plenum Press, New York, NY1994: 315
        • Marnett LJ
        • Tuttle MA
        Comparison of the mutagenicities of malondialdehyde and the side products formed during its chemical synthesis.
        Cancer Res. 1980; 40: 276
        • Parthasarathy S
        Oxidation of low-density lipoprotein by thiol compounds leads to its recognition by the acetyl LDL receptor.
        Biochim Biophys Acta. 1987; 917: 337
        • Steinbrecher UP
        • Parthasarathy S
        • Leake DS
        • Witzum JL
        • Steinberg D
        Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density lipoprotein phospholipids.
        in: 7th edn. Proc Natl Acad Sci USA. 81. 1984: 3883
        • Iuliano L
        • Pedersen JZ
        • Ghiselli A
        • Praticò D
        • Rotilio G
        • Violi F
        Mechanism of reaction of a suggested superoxide-dismutase mimic, Fe(II)-N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine.
        Arch Biochem Biophys. 1992; 293: 153
        • Halliwell B
        • Gutteridge GMC
        • Aruoma OI
        The deoxyribose method: a simple test-tube assay for determination of rate constants for reactions of hydroxyl radicals.
        Anal Biochem. 1987; 165: 215
        • Mahley RW
        • Weisgraber KH
        • Innearity TL
        • Windmueller HG
        Accelerated clearance of low-density and high-density lipoproteins and retarded clearance of E apoprotein-containing lipoproteins from the plasma of rats after modification of lysine residues.
        in: 7th edn. Proc Natl Acad Sci USA. 76. 1979: 1746
        • Fischman AJ
        • Lees AM
        • Barlai-Kovach M
        • Strauss W
        Accumulation of native and methylated low density lipoproteins by healing rabbit arterial wall.
        Arteriosclerosis. 1987; 7: 361
        • Steinbrecher UP
        • Witzum JL
        • Parthasarathy S
        • Steinberg D
        Decrease in reactive amino groups during oxidation or endothelial cell modification of LDL.
        Arterioscelrosis. 1987; 6: 135