Multicentric inflammation in epicardial coronary arteries of patients dying of acute myocardial infarction
Luigi Giusto Spagnoli, MD*,*,
Elena Bonanno, MD*,
Alessandro Mauriello, MD*,
Giampiero Palmieri, MD*,
Antonietta Partenzi, MD*,
Giuseppe Sangiorgi, MD* and
Filippo Crea, MD
* Anatomic Pathology Division, University of Rome Tor Vergata, Rome, Italy
Division of Cardiology, Catholic University, Rome, Italy
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Figure 1 Acquisition and analysis two-color flow cytometry. Panels A, B, C, D, E, and F represent the control set. (A) Monocytes from peripheral blood cells stained with CD68; the region delimits the area in which positive cells are identified. (B) Smooth muscle cells from the normal tunica media stained with alpha-smooth muscle actin (SMA); the region delimits the area in which positive cells are identified. (C) Lymphocytes from peripheral blood cells stained with CD3; the region delimits the area in which positive cells are identified. (D) Phytohemagglutinin (PHA)-activated peripheral blood cells stained with human leukocyte antigen (HLA)-DR; the region delimits the area in which positive cells are identified. (E) Coronary samples stained with control antibody (mixture of fluorescein isothiocyanate (FITC)/phycoerythrin (PE)-conjugated secondary antibodies). (F) A region gate was drawn to delimit the nonspecific stain. (G) Coronary cells stained with CD3. G1 shows single PE-immunostained cells after debris gating according to control settings. G2 shows the results in the positive region after removal of the debris. (H) Coronary cells stained with CD68. H1 shows single PE-immunostained cells after debris gating according to control settings. H2 shows the results in the positive region after removal of the debris. (I) Coronary cells stained with alpha-SMA. I1 shows single FITC-immunostained cells after debris gating according to control settings. I2 shows the results in the positive region after removal of the debris. (J) Coronary cells stained with HLA-DR. J1 shows single PE-immunostained cells after debris gating according to control settings. J2 shows the results in the positive region after removal of the debris.
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Figure 2 Distribution of cell immunophenotypes in group 1 (acute myocardial infarction [MI]), group 2 (old MI), and group 3 (no clinical history of ischemic heart disease). One-way analysis of variance showed significant differences (p = 0.0001) in the distribution of the various cytotypes and human leukocyte antigen (HLA)-DR–positive cells among the experimental groups. The Bonferroni test showed the following significant differences (p < 0.01) for actin (smooth muscle actin [SMA]) (group 1 vs. 2 and 3; group 2 vs. 3), CD3 (group 1 vs. 2 and 3; group 2 vs. 3), CD68 (group 1 vs. 2; group 2 vs. 3), and HLA-DR (group 1 vs. 2 and 3; group 2 vs. 3). Data are presented as the mean value with 95% confidence interval.
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Figure 3 Distribution of cell immunophenotypes in the infarct-related artery (IRA) and non-IRA of subjects with an acute myocardial infarction (MI) and those with an old MI. One-way analysis of variance showed significant differences (p = 0.0001) in the distribution of CD3+ (p = 0.0001), CD68+ (p = 0.0001), and human leukocyte antigen (HLA)-DR–positive cells (p = 0.004) among the experimental groups. The Bonferroni test showed the following significant differences (p < 0.05) for CD3 (a vs. b, c, and d; b vs. d; c vs. d), CD68 (a vs. c and d; b vs. d), and HLA-DR (d vs. a, b, and c). Data are presented as the mean value with 95% confidence interval.
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Figure 4 Immunohistochemical staining against human leukocyte antigen (HLA)-DR antigens and flow cytometric acquisition of HLA-DR phycoerythrin-conjugated stain in various experimental groups. (A) A coronary section from the infarct-related artery (IRA) (group 1; case no. 1 in Table 1) shows a diffuse positive reaction in the plaque and in the site of cap rupture. A1 shows the flow cytometric results of the same coronary artery after removal of the debris. (B) A coronary section from the non-IRA (same case no. 1) shows a strong positive reaction in the cells through the whole plaque. B1 shows the flow cytometric results after removal of the debris in the same coronary artery. (C) A coronary section from the IRA of a group 2 subject (case no. 22 in Table 1). The positive reaction to HLA-DR is comparable to that in the non-IRA of group 1 (see panels B and Fig. 3). C1 shows the flow cytometric results of the same coronary artery after removal of the debris. (D) A coronary section from group 3 (case no. 30 in Table 1). Only a few cells were positive for the HLA-DR antibody. D1 shows coronary cells from the same coronary artery.
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Figure 5 Confocal microscopic examination of the infarct-related artery. (A) The low power field (x100), human leukocyte antigen (HLA)-DR fluorescein isothiocyanate (FITC)-conjugated reaction shows a high number of positive cells. (B) The low power field (x100) CD3 phycoerythrin (PE)-conjugated reaction shows the presence of T lymphocytes. (C) Confocal examination of HLA-DR (FITC)/CD3 PE double staining revealed that all CD3+ cells were also positive for HLA-DR.
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