Topography of immune cell infiltration in different stages of coronary atherosclerosis revealed by multiplex immunohistochemistry

• Published in: International journal of cardiology. Heart & vasculature Vol. 44; p. 101111
• Main Authors: Cortenbach, Kimberley R.G., Morales Cano, Daniel, Meek, Jelena, Gorris, Mark A.J., Staal, Alexander H.J., Srinivas, Mangala, Jolanda M. de Vries, I., Fog Bentzon, Jacob, van Kimmenade, Roland R.J.
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier B.V 01.02.2023; Elsevier
• Subjects: Atherosclerosis; Coronary artery disease; Inflammation; Multiplex immunohistochemistry; Original Paper; mIHC; CAD; SMC; Inflammation; Coronary artery disease; ROI; LAD; EIT; MMP; Multiplex immunohistochemistry; Atherosclerosis; PIT; cDC2; PIT, pathological intimal thickening; mIHC, multiplex immunohistochemistry; CAD, coronary artery disease; LAD, left anterior descending artery; SMC, smooth muscle cell; EIT, eccentric intimal thickening; ROI, region of interest; MMP, matrix metalloproteinase; cDC2, conventional dendritic cell type 2
• ISSN: 2352-9067; 2352-9067
• DOI: 10.1016/j.ijcha.2022.101111

Overview of plaque stages and immune cells. S1/2: Shoulder 1/2. [Display omitted] Aim of this study was to investigate immune cells and subsets in different stages of human coronary artery disease with a novel multiplex immunohistochemistry (mIHC) technique. Human left anterior descending coronary artery specimens were analyzed: eccentric intimal thickening (N = 11), pathological intimal thickening (N = 10), fibroatheroma (N = 9), and fibrous plaque (N = 9). Eccentric intimal thickening was considered normal, and pathological intimal thickening, fibroatheroma, and fibrous plaque were considered diseased coronary arteries. Two mIHC panels, consisting of six and five primary antibodies, autofluoresence, and DAPI, were used to detect adaptive and innate immune cells. Via semi-automated analysis, (sub)types of immune cells in whole plaques and specific plaque regions were quantified. Increased numbers of CD3+ T cells (P < 0.001), CD20+ B cells (P = 0.013), CD68+ macrophages (P = 0.003), CD15+ neutrophils (P = 0.017), and CD31+ endothelial cells (P = 0.024) were identified in intimas of diseased coronary arteries compared to normal. Subset analyses of T cells and macrophages showed that diseased coronary arteries contained an abundance of CD3+CD8- non-cytotoxic T cells and CD68+CD206- non-M2-like macrophages. Proportions of CD3+CD45RO+ memory T cells were similar to normal coronary arteries. Among pathological intimal thickening, fibroatheroma, and fibrous plaque, all immune cell numbers and subsets were similar. The type of immune response does not differ substantially between different stages of plaque development and may provide context for mechanistic research into immune cell function in atherosclerosis. We provide the first comprehensive map of immune cell subtypes across plaque types in coronary arteries demonstrating the potential of mIHC for vascular research.