Expression of heat shock protein–70 by dendritic cells in the arterial intima and its potential significance in atherogenesis Yuri V. Bobryshev, PhD,

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Expression of heat shock protein–70 by dendritic cells in the arterial intima and its potential significance in atherogenesis Yuri V. Bobryshev, PhD, Reginald S.A. Lord, MD Journal of Vascular Surgery Volume 35, Issue 2, Pages 368-375 (February 2002) DOI: 10.1067/mva.2002.121067 Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 1 Heat shock protein–70 (HSP70) distribution in atherosclerotic plaques in carotid artery. A, Expression of HSP70 with non-foam cells (large arrows) and foam cells (small arrows) in superficial portion of atheromatous plaque. Lumen of artery is marked with asterisk. ABC immunoperoxidase technique. Visualization of HSP70 was produced with diaminobenzidine (brown-colored product). Counterstaining with Mayer's hematoxylin. B, Association of HSP70 (brown) with CD68 antigen (rose) in shoulder area of atheromatous plaque (double immunostained cells are shown with arrows ). Combination of peroxidase-antiperoxidase and alkaline phosphatase-antialkaline phosphatase immunotechniques using AEC substrate and diaminobenzidine. Counterstaining with Mayer's hematoxylin. Magnification: ×400. Journal of Vascular Surgery 2002 35, 368-375DOI: (10.1067/mva.2002.121067) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 2 Identification of heat shock protein–70 (HSP70) in dendritic cells in atherosclerotic plaque in carotid artery. A,Arrows show CD1a+ dendritic cells (rose) located on border of necrotic core in complicated atherosclerotic plaque and contained HSP70 (brown). Note intensive expression of HSP70 by foam cells surrounding necrotic (lipid) core (shown with arrows ). B,Arrows show CD1a+ dendritic cells (rose) located in deep portion of atherosclerotic intima. A,B, Combination of peroxidase-antiperoxidase and alkaline phosphatase-antialkaline phosphatase immunotechniques using AEC substrate and diaminobenzidine. Counterstaining with Mayer's hematoxylin. Magnification: ×400. Journal of Vascular Surgery 2002 35, 368-375DOI: (10.1067/mva.2002.121067) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 3 Expression of CD1d by dendritic cells in cell-rich areas in atherosclerotic plaque in carotid artery. A and B represent consecutive sections that show coexpression of CD1a (rose;A ) and CD1d (rose;B ) by dendritic cells (arrows). Immunoperoxidase technique using AEC substrate; counterstaining with Mayer's hematoxylin. C, Colocalization of CD1d+ dendritic cells (rose; arrows) with CD3+ T cells (brown). Combination of peroxidase-antiperoxidase and alkaline phosphatase-antialkaline phosphatase immunotechniques using AEC substrate and diaminobenzidine. Magnification: ×400. Journal of Vascular Surgery 2002 35, 368-375DOI: (10.1067/mva.2002.121067) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 4 Contacts of dendritic cell processes (asterisks), containing characteristic cisterns of tubulovesicular system, with lymphocyte (star;A ) and foam cell (B ). B, “Lipid droplets” are marked by stars. Electron microscopy. Magnification: ×14,000. Journal of Vascular Surgery 2002 35, 368-375DOI: (10.1067/mva.2002.121067) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 5 Association of heat shock protein–70 (HSP70) with activated dendritic cells in early intimal lesions of aorta. A, Identification of HSP70 (blue) within CD1a+ dendritic cells (rose) in intimal sheet using en face immunostaining technique. CD1a antigen was visualized with AEC substrate, and visualization of HSP70 was produced with Fast Blue substrate. Magnification: ×400. B,Arrows show cross-section of dendritic cell process, containing characteristic cisterns of tubulovesicular system, located directly under endothelium. Lumen of aorta is indicated with asterisk. Electron microscopy. Magnification: ×19,000. Journal of Vascular Surgery 2002 35, 368-375DOI: (10.1067/mva.2002.121067) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions