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. 2020 Mar 12;126(10):1346–1359. doi: 10.1161/CIRCRESAHA.119.316206

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© 2020 The Authors.

Circulation Research is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 2. — Oxidized phospholipids induce a proinflammatory EC phenotype and thereby facilitate monocyte transendothelial migration (TEM). A, Recombinant 17K apo(a) (apolipoprotein(a)) induces increased expression of TEM-associated genes (50 μg/mL; red bars). No differences were observed in ECs stimulated with 17KΔLBS (50 μg/mL; green bars). Data were analyzed using 1-way ANOVA with Tukey correction (for E-selectin, P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS; for MCP1, P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS; for ICAM1, P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS; for VCAM1, P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS; for IL6, P=0.037 for unstimulated vs 17K, P=0.0156 for 17K vs 17KΔLBS; for IL8, P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS 6 h stimulation; n=4). B, Representative differential interference contrast (DIC) images of unstimulated ECs, 17K-stimulated ECs, and 17KΔLBS-stimulated HAECs. Transmigrated monocytes are visualized as black cells with a red asterisk and adhered monocytes as white cells (18 h stimulation; n=6; white bar=200 μm). C, Quantification of adhered monocytes. Data were analyzed using 1-way ANOVA with Tukey correction. P=0.0049 for unstimulated vs 17K (17K, red bars; 17KΔLBS, green bars) and (D) transmigrated monocytes. Data were analyzed using 1-way ANOVA with Tukey correction. P<0.0001 for unstimulated vs 17K, P<0.0001 for 17K vs 17KΔLBS (18 h stimulation; n=6). E, Monoclonal antibody E06 (100 μg/mL), decreased expression of ICAM1, VCAM1, IL6, and IL8 in Lp(a) (lipoprotein(a))-ECs (green bars) vs Lp(a)-ECs (red bars). Data were analyzed using 1-way ANOVA with Tukey correction. For ICAM1, P=0.0059 for unstimulated vs Lp(a), P=0.0173 for Lp(a) vs Lp(a)+E06; for VCAM1, P=0.0187 for unstimulated vs Lp(a), P=0.0099 for Lp(a) vs Lp(a)+E06; for IL6, P=0.0162 for unstimulated vs Lp(a), P=0.0070 for Lp(a) vs Lp(a)+E06; for IL8, P=0.0324 for unstimulated vs Lp(a), P=0.0431 for Lp(a) vs Lp(a)+E06 (6 h stimulation; n=5). F, ECs incubated with 100 mg/dL Lp(a) increased monocyte adhesion (red bars) but coincubation with E06 diminished Lp(a)-induced adhesion (green bars). Data were analyzed using 1-way ANOVA with Tukey correction. P=0.0342 for unstimulated vs Lp(a), P=0.0324 for Lp(a) vs Lp(a)+E06 (18 h stimulation; n=5). G, Monocyte TEM was increased when ECs were incubated with 100 mg/dL Lp(a) (red bars) and decreased after coincubation with E06 (green bars). Data were analyzed using 1-way ANOVA with Tukey correction. P=0.0014 for unstimulated vs Lp(a), P=0.0095 for Lp(a) vs Lp(a)+E06 (18 h stimulation; n=5). All data are mean±SEM. 17K indicates 17K recombinant apolipoprotein(a); 17KΔLBS, 17K recombinant apolipoprotein(a) with a mutation in the lysine-binding site; E06, murine IgM monoclonal antibody E06 that binds the PC moiety of oxidized phospholipids; ICAM-1, intercellular adhesion molecule 1; IL, interleukin 6; MCP-1, monocyte chemoattractant protein 1; and VCAM-1, vascular adhesion molecule 1. *P<0.05, **P<0.005, ***P<0.0005, ****P<0.00005.