Cellular Immunostaining of Angiotensin-Converting Enzyme in Human Coronary Atherosclerotic Plaques
Flavio Ribichini, MD*,*,
Francesco Pugno, MD ,
Valeria Ferrero, MD*,
Gianni Bussolati, MD, FRCPath ,
Mauro Feola, MD,
Paolo Russo, MD ,
Carlo Di Mario, MD||,
Antonio Colombo, MD|| and
Corrado Vassanelli, MD*
* Catheterization Laboratory, Ospedale Maggiore della Carità, Universita’ del Piemonte Orientale, Novara, Italy
Ospedale Santa Croce e Carle, Cuneo, Italy
Dipartimento di Scienze Biomediche dell’Universita’ di Torino, Torino, Italy
Clinica Villa Maria Pia, Torino, Italy
|| Centro Cuore Columbus, Ospedale San Raffaele, Milano, Italy
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Figure 1 Double immunostaining of coronary plaque tissue from a 65-year-old man with subacute unstable angina. (A) Immunohistochemical staining (red reaction product) using anti–angiotensin I-converting enzyme antibodies and an immuno-alkaline phosphatase reaction that identifies as angiotension I-converting enzyme-positive cells the endothelium and numerous stromal cells, arranged in clusters or isolated in the stroma. (B) After removal of the red reaction product, the same area is stained in brown with an immunoperoxidase reaction revealing CD68-positive macrophages. Comparison of the two figures reveals also that some strongly angiotensin I-converting enzyme-positive cells do not correspond to macrophages.
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Figure 2 Linear regression of the association of macrophage area (mm2) identified by CD68 immunostaining and area of angiotensin I-converting enzyme-stained cells (mm2) in patients with stable angina (A) or unstable angina (B), showing both positive reactions. (A) r = 0.006, p = 0.9; (B) r = 0.6, p = 0.0001.
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Figure 3 Immunoreaction with angiotensin I-converting enzyme antibody showing positive staining of the endothelial cells of neovessels of the neointima (arrows).
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Figure 4 Double immunostaining of coronary plaque tissue from a 71-year-old woman with subacute unstable angina stained in sequence as in Figure 1. Immunohistochemical procedure for angiotensin I-converting enzyme (A: red color) and alpha-actin (B: brown color). Comparison of the two figures demonstrates that some actin-positive cells of the smooth muscle cell type are also angiotensin I-converting enzyme-positive.
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