Immunohistochemical localization of C‐reactive protein‐binding sites in human atherosclerotic aortic lesions by a modified streptavidin‐biotin‐staining method

One‐step fluorescein‐conjugated polyclonal antibody technique has shown that C‐reactive protein (CRP) was located only extracellularly in human atherosclerotic lesions. In this report a more sensitive streptavidin‐biotin technique was applied to detect the localization of CRP in human athere sclerotic lesions. lmmunohistochemical staining with polyclonal and monoclonal anti‐human CRP antibodies both produced a brown color extracellularly in the necrotic lesions, and intracelluarly in CD68+ foam cells. The latter suggests an uptake of CRP‐lipid complexes by macro‐phages. The staining is human CRP‐specific because it was eliminated by preabsorption of the monoclonal antibody with pure human CRP, or by substitution of the primary antibody with non‐immune rabbit serum. By overlaid CRP‐binding study, a positive stain was observed on intimal smooth muscle cells and foam cells, suggesting that they have CRP‐binding sites unless the CRP‐binding activity was generated de novo through the fixation procedure. Accordingly, it is hypothesized that CRP may facilitate the uptake of lipids by macrophages accumulating in atherosclerotic lesions. Further, CRP might participate in cytolysis, which enlarges the necrotic area, and/or in phagocytosis that scavenges the necrotic tissue.

[1]  H. Ishibashi-Ueda,et al.  Characterization of serum amyloid P component from human aortic atherosclerotic lesions. , 1995, Arteriosclerosis, thrombosis, and vascular biology.

[2]  A. Gown,et al.  Characterization of the Early Lesion of ‘Degenerative’ Valvular Aortic Stenosis: Histological and Immunohistochemical Studies , 1994, Circulation.

[3]  T. Blundell,et al.  Structure of pentameric human serum amyloid P component , 1994, Nature.

[4]  Y. Zhang,et al.  Plasma protein insudation as an index of early coronary atherogenesis. , 1993, The American journal of pathology.

[5]  C. Yutani,et al.  Ultrastructural studies on the phenotypic modulation of human intimal smooth muscle cells. , 1993, Atherosclerosis.

[6]  M. Jacob,et al.  Cell‐Matrix Interactions in the Genesis of Arteriosclerosis and Atheroma , 1992, Annals of the New York Academy of Sciences.

[7]  C. Mold,et al.  Serum amyloid P component binds to histones and activates the classical complement pathway. , 1992, Journal of immunology.

[8]  P. Libby,et al.  Cytokines and growth factors in atherogenesis. , 1992, Archives of pathology & laboratory medicine.

[9]  R. Christner,et al.  A cell attachment peptide from human C‐reactive protein , 1992, Journal of cellular biochemistry.

[10]  T. Ueno,et al.  Complement receptors in atherosclerotic lesions. , 1992, Artery.

[11]  N. Ku,et al.  Regulation of cytokine-induced human C-reactive protein production by transforming growth factor-beta. , 1990, Journal of immunology.

[12]  P. Butler,et al.  Pentraxin-chromatin interactions: serum amyloid P component specifically displaces H1-type histones and solubilizes native long chromatin , 1990, The Journal of experimental medicine.

[13]  I. Kushner,et al.  C-reactive protein and the acute-phase response. , 1990, Hospital practice.

[14]  N. Goldman,et al.  cis-acting elements responsible for interleukin-6 inducible C-reactive protein gene expression. , 1990, The Journal of biological chemistry.

[15]  G. Ciliberto,et al.  Dual control of C‐reactive protein gene expression by interleukin‐1 and interleukin‐6. , 1989, The EMBO journal.

[16]  G. Hansson,et al.  Complement receptors and regulatory proteins in human atherosclerotic lesions. , 1989, Arteriosclerosis.

[17]  T. D. Du Clos,et al.  C-reactive protein reacts with the U1 small nuclear ribonucleoprotein. , 1989, Journal of immunology.

[18]  G. Hansson,et al.  Prelesional complement activation in experimental atherosclerosis. Terminal C5b-9 complement deposition coincides with cholesterol accumulation in the aortic intima of hypercholesterolemic rabbits. , 1989, Laboratory investigation; a journal of technical methods and pathology.

[19]  T. D. Du Clos C-reactive protein reacts with the U1 small nuclear ribonucleoprotein. , 1989, Journal of immunology.

[20]  T. D. Du Clos,et al.  Analysis of the binding of C-reactive protein to histones and chromatin. , 1988, Journal of immunology.

[21]  Reynolds Gd,et al.  C-reactive protein immunohistochemical localization in normal and atherosclerotic human aortas. , 1987 .

[22]  R. Vance,et al.  C-reactive protein immunohistochemical localization in normal and atherosclerotic human aortas. , 1987, Archives of pathology & laboratory medicine.

[23]  G. Gabbiani,et al.  A monoclonal antibody against alpha-smooth muscle actin: a new probe for smooth muscle differentiation , 1986, The Journal of cell biology.

[24]  H. Rus,et al.  Immunoelectron-microscopic localization of the terminal C5b-9 complement complex in human atherosclerotic fibrous plaque. , 1986, Atherosclerosis.

[25]  H. Rus,et al.  Immunoglobulins and complement components in human aortic atherosclerotic intima. , 1985, Atherosclerosis.

[26]  M. Pepys,et al.  Immunohistochemical studies of c‐reactive protein and apolipoprotein b in inflammatory and arterial lesions , 1985, Journal of Pathology.

[27]  M. Pepys,et al.  Circulating human C-reactive protein binds very low density lipoproteins. , 1984, Clinical and experimental immunology.

[28]  A. Feinstein,et al.  Rabbit and rat C-reactive proteins bind apolipoprotein B-containing lipoproteins , 1984, The Journal of experimental medicine.

[29]  M. Pepys,et al.  Acute phase proteins with special reference to C-reactive protein and related proteins (pentaxins) and serum amyloid A protein. , 1983, Advances in immunology.

[30]  A. Feinstein,et al.  Low density lipoprotein and very low density lipoprotein are selectively bound by aggregated C-reactive protein , 1982, The Journal of experimental medicine.

[31]  J. Volanakis,et al.  C‐REACTIVE PROTEIN BINDING SPECIFICITIES: ARTIFICIAL AND NATURAL PHOSPHOLIPID BILAYERS * , 1982, Annals of the New York Academy of Sciences.

[32]  C. Mold,et al.  C-reactive protein and the acute phase response. , 1982, Advances in internal medicine.

[33]  R. Mortensen,et al.  Mediation of CRP-dependent phagocytosis through mouse macrophage Fc-receptors. , 1977, Journal of immunology.

[34]  T. Lint,et al.  Interaction of C-reactive protein with lymphocytes and monocytes: complement-dependent adherence and phagocytosis. , 1976, Journal of immunology.

[35]  H. Gewurz,et al.  Interactions of C-reactive protein with the complement system. III. Complement-dependent passive hemolysis initiated by CRP , 1975, The Journal of experimental medicine.

[36]  H. Gewurz,et al.  INTERACTIONS OF C-REACTIVE PROTEIN WITH THE COMPLEMENT SYSTEM , 1974, The Journal of experimental medicine.