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Increased ubiquitin immunoreactivity in unstable atherosclerotic plaques associated with acute coronary syndromes

Joerg Herrmann, MD^*, William D. Edwards, MD, David R. Holmes, Jr, MD^*, Kris L. Shogren^*, Lilach O. Lerman, MD, PhD, Aaron Ciechanover, MD, PhD and Amir Lerman, MD^*,*

^* Division of Cardiovascular Diseases, Rochester, Minnesota, USA
Division of Anatomic Pathology, Rochester, Minnesota, USA
Division of Hypertension Mayo Clinic Rochester, Rochester, Minnesota, USA
Unit of Biochemistry, Faculty of Medicine and the Rapport Institute for Research in Medical Sciences, Technion-Israel Institute of Technology, Haifa, Israel

View larger version (64K): [in a new window]   Figure 1 Illustration of the immunostaining grades applied in the current study. Original magnifications for all panels 37.5x.

View larger version (100K): [in a new window]   Figure 2 Comparative illustration of ubiquitin immunostaining in the noninfarct-related coronary artery (A) versus the infarct-related coronary artery (B). The area highlighted by the box in B is displayed at a higher magnification in C. Serial sections stained for smooth muscle alpha-actin (D) and T-cell CD 3 (E) reveal co-localization of ubiquitin immunoreactivity with inflammatory cells and neointimal cells in the cap/shoulder region. Original magnifications for A and B 15x; for C to D 100x.

View larger version (118K): [in a new window]   Figure 3 Atherosclerotic plaque area characterized by infiltration of leukocytes, most of them revealing brown staining for ubiquitin (arrow in main panel) and being identified as T cells by CD-3 immunostaining (A). Double immunofluorescence confirms co-localization of ubiquitin with T cells (round cells with yellow fluorescence for ubiquitin in the center, surrounded by red fluorescence for CD-3, B). Original magnification for all panels 100x.

View larger version (153K): [in a new window]   Figure 4 Lipid core area of an atherosclerotic plaque, in which co-localization of ubiquitin (brown) with macrophage CD 68 (red) is revealed by double-immunostaining (arrow in main panel), including nuclear staining of a foam cell (arrowhead in inserted panel). Original magnification for both panels 100x.

View larger version (117K): [in a new window]   Figure 5 Co-localization of ubiquitin (brown) with terminal deoxynucleotidyl transferase end labeling (TUNEL) positive cells (red) in lipid core area (A) and cap/shoulder region (B). Double immunofluorescence (inserted panel) more clearly visualizes ubiquitin (yellow fluorescence) in the cytoplasm of cells, which line cholesterol crystals of the lipid core and stain positive for TUNEL (red fluorescence). Original magnification for all panels 100x.

View larger version (145K): [in a new window]   Figure 6 Lipid core region (A) and cap region (B) revealing co-localization of ubiquitin (brown) with p53 (red) in macrophages (inserted panel in A, and lower inserted panel in B) and vascular smooth muscle cells (upper inserted panel in B). Original magnification for main panels and inserted panels 75x and 150x, respectively.