Monocyte subset distribution in patients with stable atherosclerosis and elevated levels of lipoprotein(a) Konstantin A. Krychtiuk, MD, Stefan P. Kastl,

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Monocyte subset distribution in patients with stable atherosclerosis and elevated levels of lipoprotein(a) Konstantin A. Krychtiuk, MD, Stefan P. Kastl, MD, Sebastian L. Hofbauer, MD, Anna Wonnerth, MD, Georg Goliasch, MD, PhD, Maria Ozsvar-Kozma, MSc, Katharina M. Katsaros, MD, Gerald Maurer, MD, Kurt Huber, MD, Elisabeth Dostal, MD, Christoph J. Binder, MD, PhD, Stefan Pfaffenberger, MD, Stanislav Oravec, MD, Johann Wojta, PhD, Walter S. Speidl, MD Journal of Clinical Lipidology Volume 9, Issue 4, Pages 533-541 (July 2015) DOI: 10.1016/j.jacl.2015.04.005 Copyright © 2015 National Lipid Association Terms and Conditions

Figure 1 Gating strategy used for monocyte subset discrimination. Monocytes were defined as CD45 positive cells (B) exhibiting a typical forward (FSC) and sideward scatter (SSC) profile (A). To exclude possible contamination with T-cells, B-cells, and natural killer cells, cells that stained for CD3, CD19, and CD56 were excluded, respectively (C). Remaining CD45+CD3/19/56– cells with a typical FSC/SSC profile were considered monocytes and distinguished according to their CD14 and CD16 surface expression into classical monocytes (CMs; CD14++CD16−), intermediate monocytes (IMs; CD14++CD16+), and nonclassical monocytes (NCMs; CD14+CD16++) (D). Journal of Clinical Lipidology 2015 9, 533-541DOI: (10.1016/j.jacl.2015.04.005) Copyright © 2015 National Lipid Association Terms and Conditions

Figure 2 Monocyte subset distribution in patients with elevated lipoprotein(a). The distribution of classical monocytes (A), intermediate monocytes (B), and non-classical monocytes (C) in patients with normal (≤50 mg/dL) and elevated lipoprotein(a) (>50 mg/dL) were determined as described under Methods. Bar graphs indicate mean % of total monocytes and error bars represent standard deviation. *P < .05. Journal of Clinical Lipidology 2015 9, 533-541DOI: (10.1016/j.jacl.2015.04.005) Copyright © 2015 National Lipid Association Terms and Conditions

Figure 3 Association of oxidized phospholipids with lipoprotein(a) levels and intermediate monocytes (IMs). Plasma levels of OxPLP were determined as described under Methods in patients with normal (≤50 mg/dL) and elevated lipoprotein(a) (>50 mg/dL) (A). Correlation of oxidized phospholipid (OxPLP) with IM (B). *P < .0001. Journal of Clinical Lipidology 2015 9, 533-541DOI: (10.1016/j.jacl.2015.04.005) Copyright © 2015 National Lipid Association Terms and Conditions

Figure 4 Circulating inflammatory markers in patients with normal and elevated lipoprotein(a). Plasma levels of C-reactive protein (log-scale) (A), interleukin (IL)-6 (log-scale) (B), and IL-10 (C) were determined as described under Methods in patients with normal (≤50 mg/dL) and elevated lipoprotein(a) (>50 mg/dL). Box plots represent median, interquartile, and total range. *P < .05. Journal of Clinical Lipidology 2015 9, 533-541DOI: (10.1016/j.jacl.2015.04.005) Copyright © 2015 National Lipid Association Terms and Conditions