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Biochemical and Immunological Evidence for the Presence of Lipoprotein-Antilipoprotein Immune Complexes in Human Atherosclerotic Plaques

  • William Hollander
  • Antonio Lazzari
  • Carl Franzblau
Conference paper

Summary

Previous immunohistochemical studies of human atherosclerotic plaques have indicated a close association of the lipid protein complexes with the immunoglobulins (IgA, IgG, IgM) and complement (C3) contained in the lesion. The aim of the present studies was to determine whether the lipid-protein complexes or altered lipoproteins contained in the atherosclerotic plaque are present in the form of lipoprotein-antibody immune complexes.

The lipid-protein complexes were extracted from aortic atherosclerotic plaques into phosphate buffered saline and separated in the ultracentrifuge into a very low density fraction of d < 1.006 g/mL (VLDF) and a low density fraction of d 1.063–1.006 g/mL (LDF). LDF and VLDF were then purified by gel filtration on a Bio-Gel A-150 m column. The fractions were rich in cholesteryl ester and contained immunoreactive apo A-I, B, C-III and E as well as IgG, IgM, and C3. Small amounts of glycosaminoglycans (GAGs) consisting mainly of hyaluronic acid and chondroitin sulfate-C were detected in the fractions as was calcium. When examined by scanning electron microscope LDF and VLDF appeared heterogeneous in size and shape and showed surface irregularities and defects consistent with the appearance of aggregated and degraded lipoproteins. When mouse peritoneal macrophages were incubated with LDF or VLDF, they accumulated cholesterol and cholesteryl ester and became foam cells. In contrast to the macrophages, smooth muscle cells did not accumulate lipid when incubated with LDF or VLDF. The uptake of the lipid-protein complexes by the macrophage appeared to be mediated by the protein component of the complexes since pretreatment of LDF or VLDF with pronase completely inhibited lipid deposition in the macrophage. Digestion of LDF or VLDF with chondroitinase ABC and hyaluronidase removed the GAG components of the complexes but it did not reduce lipid accumulation in the macrophage suggesting that the uptake of the complexes is not mediated by the dextran sulfate receptors of the macrophage. Competitive inhibition studies with opsonized sheep red blood cells suggest that the immune receptors (Fc and C3b receptors) of the macrophage may mediate the uptake of LDF and VLDF in a manner similar to that of an immune complex. The protein composition of foam cells isolated from atherosclerotic plaques, as revealed by immunohistochemical examination, appeared to be similar to that of the lipid-protein complexes and mouse peritoneal macrophages pre-incubated with these complexes suggesting that the complexes may play a role in the formation of macrophage derived foam cells in atherosclerotic lesions. The overall results of the study suggest that the lipid-protein complexes, which account for over 90% of the lipoprotein fractions isolated from atherosclerotic lesions, are altered plasma lipoproteins which have the biochemical and biological behavior of a lipoprotein-antibody immune complex.

Keywords

Atherosclerotic Plaque Cholesteryl Ester Peritoneal Macrophage Foam Cell Mouse Peritoneal Macrophage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Hollander W, Colombo MA, Kramsch DM, and Kirkpatrick BJ (1974) Immunological aspects of atherosclerosis. Comparative pathology of the heart. Adv Cardiol 13: 192–207.PubMedGoogle Scholar
  2. 2.
    Hollander W, Colombo MA, Kirkpatrick BJ, Paddock J (1979) Soluble proteins in the human atherosclerotic plaque: With special reference to immunoglobulins, C3 complement component, α1-antitrypsin and α2-macroglobulin. Atherosclerosis 34:391–405.PubMedCrossRefGoogle Scholar
  3. 3.
    Parums D, Mitchinson MJ (1981) Demonstration of immunoglobulin in the neighborhood of advanced atherosclerotic plaques. Atherosclerosis 38:211–216.PubMedCrossRefGoogle Scholar
  4. 4.
    Hansson GK, Holm J, Kral JG (1984) Accumulation of IgG and complement factor C3 in human arterial endothelium and atherosclerotic lesions. Acta Pathol Microbiol Immunol Scand 92:429–435.Google Scholar
  5. 5.
    Vlaicu R, Rus HG, Niculescu F, Cristea A (1985) Immunoglobulins and complement components in human aortic atherosclerotic intima. Atherosclerosis 55:35–50.PubMedCrossRefGoogle Scholar
  6. 6.
    Rus HG, Niculescu F, Constantinescu E, Cristea A, Vlaicu R (1986) Immunoelectron-microscopic localization of the terminal C5b-9 complement complex in human atherosclerotic fibrous plaque. Atherosclerosis 6: 35–42.CrossRefGoogle Scholar
  7. 7.
    Schwartz CJ, Mitchell JRA (1962) Cellular infiltration of the human arterial adventitia associated with atheromatous plaques. Circulation 26:73–78.PubMedGoogle Scholar
  8. 8.
    Stratford N, Britten K, Gallagher P (1986) Inflammatory infiltrates in human coronary atherosclerosis. Atherosclerosis 59:271–276.PubMedCrossRefGoogle Scholar
  9. 9.
    Jonasson L, Holm J, Skalli O, Bondjers G, Hansson GK (1986) Regional accumulations of T cells, macrophages and smooth muscle cells in the human atherosclerotic plaque. Arteriosclerosis 6:131–138.PubMedCrossRefGoogle Scholar
  10. 10.
    Beaumont JL (1970) Autoimmune hyperlipidemia. In: Jones RJ (ed) Atherosclerosis. Berlin: Springer-Verlag, pp 166–176.Google Scholar
  11. 11.
    Szondy E, Horvath M, Mezey Z, Szekely J, Lengyel E, Fust G, Gero S (1983) Free and complexed anti-lipoprotein antibodies in vascular diseases. Atherosclerosis 49:67–77.CrossRefGoogle Scholar
  12. 12.
    Klimov AN, Denisenko AD, Popov AV, Nagornev VA, Pleskov VM, Vinogradov AG, Densisenko TV, Magracheva EY, Kheifes GM, Kuznetzov AS (1985) Lipoprotein-antibody immune complexes: Their catabolism and role in foam cell formation. Atherosclerosis 58:1–15.PubMedCrossRefGoogle Scholar
  13. 13.
    Romano EL, Camejo G, Soyano A (1984) Circulating immune complexes and antibodies to dietary antigens in patients with occlusive coronary lesions. Atherosclerosis 53:119.PubMedCrossRefGoogle Scholar
  14. 14.
    Shingu M, Hurd ER (1981) Sera from patients with systemic lupus erythematosus reactive with human endothelial cells. J Rheumatol 8:581.PubMedGoogle Scholar
  15. 15.
    Caplan M, Hastillo A, Hess ML (1984) Immunologic mechanism in atherosclerotic process. Cardiovas Rev 5:713.Google Scholar
  16. 16.
    Preston RN (1979) Immunological aspects of atherosclerosis: A review. J Royal Soc Med 72:674.Google Scholar
  17. 17.
    Hollander W, Paddock J, Colombo MA (1979) Lipoproteins in human atherosclerotic vessels. I. Biochemical properties of arterial low density lipoproteins, very low density lipoproteins, and high density lipoproteins. Exp Mol Pathol 30:144–171.PubMedCrossRefGoogle Scholar
  18. 18.
    Hollander W, Paddock J, Colombo MA (1979) Lipoproteins in human atherosclerotic vessels. II. Biochemical properties of the major apolipoproteins of arterial low density and very low density lipoproteins. Exp Mol Pathol 30:172–189.PubMedCrossRefGoogle Scholar
  19. 19.
    Ball RY, Bindman JP, Carpenter KLH, Mitchinson MJ (1986) Oxidized low density lipoprotein induces caroid accumulation by murine peritoneal macrophages in vitro. Atherosclerosis 60:173–181.PubMedCrossRefGoogle Scholar
  20. 20.
    Havel RJ, Eder HA, Bragdon JH (1955) The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J Clin Invest 34: 1345–1353.PubMedCrossRefGoogle Scholar
  21. 21.
    Schoenheimer R, Sperry WM (1934) A micromethod for the determination of free and combined cholesterol. J Biol Chem 106:745–760.Google Scholar
  22. 22.
    Youngberg GE, Youngberg MV (1930) Phosphorus metabolism. I. A system of blood phosphorus analysis. J Lab Clin Med 16:158–166.Google Scholar
  23. 23.
    Van Handel E, Zilversmit DB (1957) Micromethod for the direct determination of serum triglycerides. J Lab Clin Med 50:152.Google Scholar
  24. 24.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin reagent. J Biol Chem 193:265–275.PubMedGoogle Scholar
  25. 25.
    Metcalfe LD, Schmitz AA (1961) The rapid preparation of fatty acid esters for gas chromatographic analysis. Anal Chem 33:363–364.CrossRefGoogle Scholar
  26. 26.
    Stevens RL, Colombo MA, Gonzales JJ, Hollander W, Schmid K (1976) The glycosaminoglycans of the human artery and their changes in atherosclerosis. J Clin Invest 58:470.PubMedCrossRefGoogle Scholar
  27. 27.
    Hata R, Nagai Y (1973) A micro colorometric determination of acidic glycosaminoglycans by two-dimensional electrophoresis on a cellulose acetate strip. Anal Biochem 52:652.PubMedCrossRefGoogle Scholar
  28. 28.
    Noble RP (1968) Electrophoretic separation of plasma lipoproteins in agarose gel. J Lipid Res 9:693.PubMedGoogle Scholar
  29. 29.
    Weber D, Osborn M (1969) The reliability of molecular weight determinations by duodecyl-sulphate polyacryl-amide gel electrophoresis. J Biol Chem 244:4405–4412.Google Scholar
  30. 30.
    Ouchterlony O (1958) Diffusion-in-gel methods for immunological analysis. Progr Allergy 5:1–78.Google Scholar
  31. 31.
    Scheidegger JJ (1955) Une micro-method de l’ immunoelectrophorese. Int Arch Allergy 7:103–110.PubMedCrossRefGoogle Scholar
  32. 32.
    Insull W Jr, Hata Y (1971) Morphology of lipid rich organelles in tissues of man and rat. Proc IV Annual Scanning Electron Microscope Symposium, pp 337–341.Google Scholar
  33. 33.
    Edelson PJ, Cohn ZA (1976) Purification and cultivation of monocytes and macrophages. In: Bloom BR, David JR (eds) In vitro Methods in Cell-Mediated and Tumor Immunity. New York: Academic Press, Inc, pp 333–340.Google Scholar
  34. 34.
    Oakes BW, Batty AC, Handley CJ, Sandberg LB (1982) The synthesis of elastin, collagen, glycosaminoglycans by high density primary cultures of neonatal rat aortic smooth muscle: An ultrastructural and biochemical study. European J Cell 27:34–46.Google Scholar
  35. 35.
    Schaffner T, Taylor K., Burtucci EJ, Fisher-Dzoga K, Beeson JH, Glagov S, Wissler RW (1980) Arterial foam cells with distinctive immunomorphological and histochemical features of macrophages. Amer J Pathol 100:57.Google Scholar
  36. 36.
    Gamble W, Vaughn M, Avigan J (1978) Procedure for determination of free and total cholesterol in cultured cells. J Lipid Res 19:1068.PubMedGoogle Scholar
  37. 37.
    Griffin FM, Silverstein SC (1974) Segment response of the macrophage plasma membrane to a phagocytic stimulus. J Exp Med 139:323.PubMedCrossRefGoogle Scholar
  38. 38.
    Coons AH, Kaplan MH (1950) Localization of antigen in tissue cells, Part 2. Improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med 91:1PubMedCrossRefGoogle Scholar
  39. 39.
    Srinivasan SR, Dolan P, Radhakrishnamurtry B, Berenson GS (1972) Isolation of lipoprotein-acid-mucopoly-saccharide complexes from fatty streaks of human aorta. Atherosclerosis 16:95–104.PubMedCrossRefGoogle Scholar
  40. 40.
    Smith EB, Evans PH, Downham MD (1967) Lipid in the aortic intima: The correlation of morphological and chemical characteristics. J Atheroscler Res 7:171–186.PubMedCrossRefGoogle Scholar
  41. 41.
    Steinbrecher UP, Fischer M, Witztum JL, Curtiss LK (198 4) Immunogenicity of homologous low density lipoprotein after methylation, ethylation, acetylation and carbamylation. Generation of antibodies specific for derived lysine. J Lipid Res 24:1109.Google Scholar
  42. 42.
    Basu S, Brown MS, Ho YK, Goldstein JL (1979) Degradation of low density lipoprotein-dextran sulfate complexes associated with deposition of cholesteryl esters in mouse macrophages. J Biol Chem 254:7141.PubMedGoogle Scholar
  43. 43.
    Goldstein JL, Hoff HF, Yo YK, Basu S, Brown MS (1981) Stimulation of cholesteryl ester synthesis in macrophages by extracts of atherosclerotic human aortas and complexes of albumin/cholesteryl esters. Arteriosclerosis 1:210.PubMedCrossRefGoogle Scholar
  44. 44.
    Yomantas S, Elner VM, Schaffner T, Wissler RW (1984) Immunohistochemical localization of apolipoprotein B in human atherosclerotic lesions. Arch Pathol Lab Med 108:374–378.PubMedGoogle Scholar
  45. 45.
    Murase T, Oka T, Yamada N, Mori N, Ishibashi S, Takaku F, Mori W (1986) Immunohistochemical localization of apolipoprotein E in atherosclerotic lesions of the aorta and coronary arteries. Atherosclerosis 60:1–6.PubMedCrossRefGoogle Scholar
  46. 46.
    Nathan CF, Murray HW, Cohn ZA (1980) The macrophage as an effector cell. New Engl J Med 303:622–626.PubMedCrossRefGoogle Scholar
  47. 47.
    Basu SK, Ho YK, Brown MS, Bilheimer DW, Anderson RW, Goldstein JL (1982) Biochemical and genetic studies of the apoprotein E secreted by mouse macrophages and human monocytes. J Biol Chem 257:9788–9795.PubMedGoogle Scholar
  48. 48.
    Werb Z, Chin JR (1983) Apoprotein E is synthesized by resident and thioglycollate-elicited macrophages but not by pyran copolymer of bacillus Calmette-Guerin activated macrophages. J Exp Med 158:1271–1293.CrossRefGoogle Scholar
  49. 49.
    Driscoll DM, Getz GS (1984) Extrahepatic synthesis of apolipoprotein E. J Lipid Res 25:1368–1379.Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1990

Authors and Affiliations

  • William Hollander
  • Antonio Lazzari
  • Carl Franzblau

There are no affiliations available

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