Advertisement

Journal of Nuclear Cardiology

, Volume 6, Issue 1, pp 41–53 | Cite as

Radiolabeled MDA2, an oxidation-specific, monoclonal antibody, identifies native atherosclerotic lesions in vivo

  • Sotirios TsimikasEmail author
  • Wulf Palinski
  • Samuel E. Halpern
  • David W. Yeung
  • Linda K. Curtiss
  • Joseph L. Witztum
Original Articles

Abstract

Background

Oxidatively modified low-density lipoprotein (LDL) is present in atherosclerotic but not normal arteries and plays a crucial role in the pathogenesis and adverse consequences of atherosclerotic lesions. We previously generated a series of monoclonal antibodies (MoAb) against oxidation-specific neo-epitopes formed during the oxidative modification of LDL. MDA2, a prototype MoAb, recognizes malondialdehyde-lysine epitopes (eg, in malondialdehyde-modified LDL) within atherosclerotic lesions. We describe the in vivo characteristics of MDA2 and initial noninvasive imaging studies of atherosclerosis in rabbits.

Methods

To assess the in vivo specificity of MDA2 for atherosclerotic lesions, iodine 125-MDA2 was intravenously injected into 7 LDL-receptor deficient Watanabe heritable hyperlipidemic (WHHL) and 2 normal New Zealand white (NZW) rabbits, and the aortic plaque uptake was evaluated 24 hours later. 125I-Halb, an isotype-matched irrelevant MoAb that binds to human albumin, was injected into 5 WHHL and 2 NZW rabbits as a control. Aortic autoradiography was performed, and the mean uptake of MoAbs was measured as the percent injected dose per gram aortic tissue. Gamma camera imaging was then carried out in 7 WHHL rabbits and 2 NZW rabbits with 99mTc-MDA2. Imaging was carried out at 10 minutes and at 12 or 24 hours. Malondialdehyde-LDL was then injected to clear the blood pool signal, and final images were obtained 2 hours later.

Results

Mean uptake of 125I-MDA2 in the entire aorta was 17.4-fold higher in WHHL than in NZW aortas (P<.001), and 2.8-fold higher than 125I-Halb in WHHL aortas. 125I-MDA2 also had higher specificity for lesioned areas than 125I-Halb (plaque/normal ratio 6.3 vs 2.9, P<.001). Autoradiograph of aortas of 125I-MDA2-injected WHHL rabbits revealed uptake in lipid-stained lesions with absence of signal in adjacent normal arterial tissue. Immunostaining of WHHL lesions, which accumulated MDA2 as noted on autoradiography, revealed that uptake was highest in areas with abundant foam cells and in lipid-rich necrotic core areas. Autoradiograph of aortas from NZW rabbits injected with 125I-MDA2 did not yield any visible signal. Planar gamma camera in vivo scintigraphy revealed a visible signal in 4/7 WHHL rabbits, which was confirmed by aortic Sudan staining.

Conclusion

Radiolabeled MDA2 shows excellent in vivo uptake and specificity for atherosclerotic lesions containing abundant oxidation-specific epitopes. The in vivo imaging studies suggest that noninvasive imaging of oxidation-rich atherosclerotic lesions with radiolabeled MDA2 may be feasible in human beings with optimization of the imaging methods.

Key Words

arteriosclerosis imaging antibodies oxidation oxidized LDL radioisotopes 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    White CW, Wright CB, Doty DB, et al. Does visual interpretation of the coronary angiogram predict physiologic significance of a coronary stenosis? N Engl J Med 1984;310:819–24.PubMedGoogle Scholar
  2. 2.
    Meier B, Gruentzig AR, Goebel N, Pyle R, von Gosslar W, Schlumf M. Assessment of stenoses in coronary angioplasty: inter- and intraobserver variability. Int J Cardiol 1983;3:159–69.PubMedCrossRefGoogle Scholar
  3. 3.
    Stiehl G, Ludmilla SG, Schofer J, Donath K, Mathey DG. Impact of compensatory enlargement of atherosclerotic coronary arteries on angiographic assessment of coronary artery disease. Circulation 1989;80:1603–9.Google Scholar
  4. 4.
    Glagov S, Weisenberg E, Zarins CK, Stankunavicius R, Kolettis GJ. Compensatory enlargement of human atherosclerotic coronary arteries. N Engl J Med 1987;316:1371–5.PubMedGoogle Scholar
  5. 5.
    Furberg CD, Adams HP, Applegate WB, et al. Effect of lovastatin on early carotid atherosclerosis and cardiovascular events. Circulation 1994;90:1679–87.PubMedGoogle Scholar
  6. 6.
    Yock PG, Johnson EL, Linker DT. Intravascular ultrasound: development and clinical potential. Am J Card Imaging 1988;2:185–93.Google Scholar
  7. 7.
    Wong ND, Detrano RC, Abrahamson D, Tobis JM, Gardin JM. Coronary artery screening by electron beam computed tomography: facts, controversy, and future. Circulation 1995;92:632–6.PubMedGoogle Scholar
  8. 8.
    Yuan C, Skinner MP, Kaneko E, et al. Magnetic resonance imaging to study lesions of atherosclerosis in the hyperlipidemic rabbit aorta. Magn Reson Imaging 1996;14:93–102.PubMedCrossRefGoogle Scholar
  9. 9.
    Constantinides P. Plaque fissures in human coronary thrombosis. J Atheroscler Res 1966;6:1–17.CrossRefGoogle Scholar
  10. 10.
    Davies MJ, Thomas A. Thrombosis and acute coronary-artery lesions in sudden cardiac ischemic death. N Engl J Med 1984;310:1137–40.PubMedGoogle Scholar
  11. 11.
    Little WC, Constantinescu M, Applegate RJ, et al. Can coronary angiography predict the site of a subsequent myocardial infarction in patients with mild-to-moderate coronary artery disease? Circulation 1988;78:1157–66.PubMedGoogle Scholar
  12. 12.
    Hackett D, Davies MJ, Maseri A. Pre-existing coronary stenoses in patients with first myocardial infarction are not necessarily severe. Eur Heart J 1988;9:1317–23.PubMedGoogle Scholar
  13. 13.
    Lendon CL, Davies MJ, Born GVR, Richardson PD. Atherosclerotic plaque caps are locally weakened when macrophages density is increased. Atherosclerosis 1991;87:87–90.PubMedCrossRefGoogle Scholar
  14. 14.
    van der Wal AC, Becker AE, van der Loos CM, Das PK. Site of intimal rupture or erosion of thrombosed coronary atherosclerotic plaques is characterized by an inflammatory process irrespective of the dominant plaque morphology. Circulation 1994;89:36–44.PubMedGoogle Scholar
  15. 15.
    Libby P. Molecular bases of the acute coronary syndromes. Circulation 1995;91:2844–50.PubMedGoogle Scholar
  16. 16.
    Shah PK, Falk E, Badimon JJ, et al. Human monocyte-derived macrophages induce collagen breakdown in fibrous caps of atherosclerotic plaques. Circulation 1995;92:1565–9.PubMedGoogle Scholar
  17. 17.
    Lees RS, Lees AM, Strauss HW. External imaging of human atherosclerosis. J Nucl Med 1983;24:154–6.PubMedGoogle Scholar
  18. 18.
    Lees AM, Lees RS, Schoen FJ, et al. Imaging human atherosclerosis with Tc-99m-labeled LDL. Arterioscler Thromb 1988;8:461–70.Google Scholar
  19. 19.
    Rosen JM, Butler SP, Meineken GE, et al. Indium-111-labeled LDL: a potential agent for imaging atherosclerotic disease and lipoprotein biodistribution. J Nucl Med 1990;31:343–50.PubMedGoogle Scholar
  20. 20.
    Hardoff R, Braegelman F, Zanzonico P, et al. External imaging of atherosclerosis in rabbits using an 123I-labeled synthetic peptide fragment. J Clin Pharmacol 1993;33:1039–47.PubMedGoogle Scholar
  21. 21.
    Davis HH II, Siegel BA, Sherman LA, et al. Scintigraphic detection of carotid atherosclerosis with indium-111-labeled autologous platelets. Circulation 1980;61:982–8.PubMedGoogle Scholar
  22. 22.
    Minar E, Ehringer H, Dudczak R, et al. Indium-111-labeled platelet scintigraphy in carotid atherosclerosis. Stroke 1989;20:27–33.PubMedGoogle Scholar
  23. 23.
    Moriwaki H, Matsumoto M, Handa N, et al. Functional and anatomic evaluation of carotid atherothrombosis: a combined study of indium 111 platelet scintigraphy and B-mode ultrasonography. Arterioscler Thromb Vasc Biol 1995;15:2234–40.PubMedGoogle Scholar
  24. 24.
    Miller DD, Boulet AJ, Tio FO, et al. In vivo technetium-99m S12 antibody imaging of platelet alpha-granules in rabbit endothelial neointimal proliferation after angioplasty. Circulation 1991;83:224–36.PubMedGoogle Scholar
  25. 25.
    Fischman AJ, Rubin RH, Khaw BA, et al. Radionuclide imaging of experimental atherosclerosis with nonspecific polyclonal immunoglobulin G. J Nucl Med 1989;30:1095–100.PubMedGoogle Scholar
  26. 26.
    Demacker PNM, Dormans TPJ, Koenders EB, Corstens FHM. Evaluation of indium-111-polyclonal immunoglobulin G to quantitate atherosclerosis in Watanabe heritable hyperlipidemic rabbits with scintigraphy: effect of age and treatment with antioxidants or ethinylestradiol. J Nucl Med 1993;34:1316–21.PubMedGoogle Scholar
  27. 27.
    Palac RT, Gray LL, Turner FE, Brown PH, Malinow MR, Demots H. Detection of experimental atherosclerosis with indium-111 radiolabeled hematoporphyrin derivative. Nucl Med Comm 1989;10:841–50.CrossRefGoogle Scholar
  28. 28.
    Chakrabarti M, Cheng KT, Spicer KM, et al. Biodistribution and radioimmunopharmacokinetics of 131I-Ama MoAb in atherosclerotic rabbits. Nucl Med Biol 1995;22:693–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Narula J, Petrov A, Bianchi C, et al. Noninvasive localization of experimental atherosclerotic lesions with mouse/human chimeric Z2D3 F(ab′)2 specific for the proliferating smooth muscle cells of human atheroma: imaging with conventional and negative charge-modified antibody fragments. Circulation 1995;92:474–84.PubMedGoogle Scholar
  30. 30.
    Palinski W, Ylä-Herttuala S, Rosenfeld ME, et al. Antisera and monoclonal antibodies specific for epitopes generated during oxidative modification of low density lipoprotein. Arterioscler Thromb 1990;10:325–35.Google Scholar
  31. 31.
    Palinski W, Rosenfeld ME, Ylä-Herttuala S, et al. Low density lipoprotein undergoes oxidative modification in vivo. Proc Natl Acad Sci 1989;86:1372–6.PubMedCrossRefGoogle Scholar
  32. 32.
    Palinski W, Hörkkö S, Miller E, et al. Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice: demonstration of epitopes of oxidized low density lipoprotein in human plasma. J Clin Invest 1996;98:800–14.PubMedCrossRefGoogle Scholar
  33. 33.
    Steinbrecher UP. Oxidation of human low density lipoprotein results in derivatization of lysine residues of apolipoprotein B by lipid peroxide decomposition products. J Biol Chem 1987;262:3603–8.PubMedGoogle Scholar
  34. 34.
    Steinberg D, Parthasarathy S, Carew TE, Khoo JC, Witztum JL. Beyond cholesterol. Modifications of low-density lipoprotein that increase its atherogenicity. N Engl J Med 1989;320:915–24.PubMedGoogle Scholar
  35. 35.
    Rosenfeld ME, Palinski W, Ylä-Herttuala S, Butler SW, Witztum JL. Distribution of oxidation-specific lipid-protein adducts and apolipoprotein B in atherosclerotic lesions of varying severity from WHHL rabbits. Arterioscler Thromb 1990;10:336–49.Google Scholar
  36. 36.
    Palinski W, Miller E, Witztum JL. Immunization of low density lipoprotein (LDL) receptor-deficient rabbits with homologous malondialdehyde-modified LDL reduces atherogenesis. Proc Natl Acad Sci 1995;92:821–5.PubMedCrossRefGoogle Scholar
  37. 37.
    Boyd HC, Gown AM, Wolfbauer G, Chait A. Direct evidence for a protein recognized by a MoAb against oxidatively modified LDL in atherosclerotic lesions from a Watanabe heritable hyperlipidemic rabbit. Am J Pathol 1989;135:815–25.PubMedGoogle Scholar
  38. 38.
    Haberland ME, Fong D, Cheng L. Malondialdehyde-altered protein occurs in atheroma of Watanabe heritable hyperlipidemic rabbits. Science 1988;241:215–8.PubMedCrossRefGoogle Scholar
  39. 39.
    Palinski W, Tangirala RK, Miller E, Young SG, Witztum JL. Increased autoantibody titers against epitopes of oxidized LDL in LDL receptor-deficient mice with increased atherosclerosis. Arterioscler Thromb Vasc Biol 1995;15:1569–76.PubMedGoogle Scholar
  40. 40.
    Palinski W, Ord VA, Plump AS, Breslow JL, Steinberg D, Witztum JL. ApoE-deficient mice are a model of lipoprotein oxidation in atherogenesis: demonstration of oxidation-specific epitopes in lesions and high titers of autoantibodies to malondialdehyde-lysine in serum. Arterioscler Thromb 1994;14:605–16.PubMedGoogle Scholar
  41. 41.
    Ylä-Herttuala S, Palinski W, Rosenfeld ME, et al. Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man. J Clin Invest 1989;84:1086–95.PubMedCrossRefGoogle Scholar
  42. 42.
    Jürgens G, Chen Q, Esterbauer H, Mair S, Ledinski G, Dinges HP. Immunostaining of human autopsy aortas with antibodies to modified apolipoprotein B and apoprotein(a). Arterioscler Thromb 1993;13:1689–99.PubMedGoogle Scholar
  43. 43.
    Hammer A, Kager G, Dohr G, Rabl H, Ghassempur I, Jürgens G. Generation, characterization, and histochemical application of monoclonal antibodies selectively recognizing oxidatively modified apoB-containing serum lipoproteins. Arterioscler Thromb Vasc Biol 1995;15:704–13.PubMedGoogle Scholar
  44. 44.
    Jue R, Lambert JM, Pierce LR, Traut RR. Addition of sulfhydryl groups to Escherichia coli ribosomes by protein modification with 2-iminothialane (methyl 4-mercaptobutyrimidate). Biochemistry 1978;17:5399–416.PubMedCrossRefGoogle Scholar
  45. 45.
    Hörkkö S, Miller E, Dudl E, et al. Antiphospholipid antibodies are directed against epitopes of oxidized phospholipids. Recognition of cardiolipin by monoclonal antibodies to epitopes of oxidized low density lipoprotein. J Clin Invest 1996;98:815–25.PubMedCrossRefGoogle Scholar
  46. 46.
    Habeeb ASSA. Chemical evaluation of conformational differences in native and chemically modified proteins by trinitrobenzenesulfonic acid. Biochim Biophys Acta 1966;115:440–54.PubMedGoogle Scholar
  47. 47.
    Fruebis J, Carew TE, Palinski W. Effect of vitamin E on atherogenesis in LDL receptor-deficient rabbits. Arterioscler Thromb Vasc Biol 1995;117:217–24.Google Scholar
  48. 48.
    Tangirala RK, Rubin EM, Palinski W. Quantitation of atherosclerosis in murine models: correlation between lesions in the aortic origin and in the entire aorta, and differences in the extent of lesions between sexes in LDL receptor-deficient and apoprotein E-deficient mice. J Lipid Res 1995;36:2320–8.PubMedGoogle Scholar
  49. 49.
    Palinski W, Koschinsky T, Butler SW, et al. Immunological evidence for the presence of advanced glycosylation end products in atherosclerotic lesions of euglycemic rabbits. Arterioscler Thromb Vasc Biol 1995;15:571–82.PubMedGoogle Scholar
  50. 50.
    Adams CWM, Virag S, Morgan RS, Orton CC. Dissociation of [3H]cholesterol and 125I-labeled plasma protein influx in normal and atheromatous rabbit aorta. J Atheroscler Res 1968;8:679–96.PubMedCrossRefGoogle Scholar
  51. 51.
    Christensen S, Nielsen H. Permeability of arterial endothelium to plasma macromolecules. Atherosclerosis 1977;27:447–63.PubMedCrossRefGoogle Scholar
  52. 52.
    Smith EB, Staples EM. Distribution of plasma proteins across the human aortic wall. Atherosclerosis 1980;37:579–90.PubMedCrossRefGoogle Scholar
  53. 53.
    Jensen JS. Renal and systemic transvascular albumin leakage in severe atherosclerosis. Arterioscler Thromb Vasc Biol 1995;15:1324–9.PubMedGoogle Scholar
  54. 54.
    Wu CC, Chang SW, Chen MS, Lee YT. Early change of vascular permeability in hypercholesterolemic rabbits. Arterioscler Thromb Vasc Biol 1995;15:529–33.PubMedGoogle Scholar
  55. 55.
    Witztum JL. Role of oxidised low density lipoprotein in atherogenesis. Br Heart J 1993;69:S12-S18.PubMedCrossRefGoogle Scholar
  56. 56.
    Scanlon CEO, Berger B, Malcom G, Wissler RW. Evidence for more extensive deposits of epitopes of oxidized low density lipoprotein in aortas of young people with elevated serum thiocyanate levels. Atherosclerosis 1996;121:23–33.PubMedCrossRefGoogle Scholar
  57. 57.
    Rosenfeld ME, Tsukada T, Chait A, Bierman EL, Gown AM, Ross R. Fatty streak expansion and maturation in Watanabe heritable hyperlipemic and comparably hypercholesterolemic fat-fed rabbits. Arteriosclerosis 1987;7:24–34.PubMedGoogle Scholar
  58. 58.
    Sinitsyn W, Mamontova AG, Checkneva W, Shnyra AA, Domogatsky SP. Rapid blood clearance of biotinylated IgG after infusion of avidin. J Nucl Med 1989;30:66–9.PubMedGoogle Scholar
  59. 59.
    Paganelli G, Malcovati M, Fazio F. Monoclonal antibody pretargetting techniques for tumour localization: the avidin-biotin system. Nucl Med Comm 1991;12:211–34.CrossRefGoogle Scholar

Copyright information

© American Society of Nuclear Cardiology 1999

Authors and Affiliations

  • Sotirios Tsimikas
    • 1
    Email author
  • Wulf Palinski
    • 2
  • Samuel E. Halpern
    • 3
  • David W. Yeung
    • 3
    • 4
  • Linda K. Curtiss
    • 5
  • Joseph L. Witztum
    • 2
  1. 1.Department of Medicine, Division of Cardiovascular DiseaseUniversity of California, San DiegoLa Jolla
  2. 2.Endocrinology and MetabolismUniversity of California, San DiegoLa Jolla
  3. 3.Department of Radiology of the Veterans Administration Medical CenterUniversity of California, San DiegoLa Jolla
  4. 4.Nuclear Medicine Service of the Veterans Administration Medical CenterUniversity of California, San DiegoLa Jolla
  5. 5.Department of Immunology and Vascular BiologyThe Scripps Research InstituteLa Jolla

Personalised recommendations