Muffova Barbora, Kauerova Sona, Bartuskova Hana, Paukner Karel, Janousek Libor, Cermakova Helena, Fronek Jiri, Kollar Marek, Petras Marek, Pitha Jan, Poledne Rudolf, Kralova Lesna Ivana
Laboratory for Atherosclerosis Research, Centre for Experimental Medicine Institute for Clinical and Experimental Medicine Prague Czech Republic.
Department of Physiology, Faculty of Science Charles University Prague Czech Republic.
FASEB Bioadv. 2025 May 5;7(6):e70017. doi: 10.1096/fba.2025-00037. eCollection 2025 Jun.
The importance of macrophage polarization through atherogenesis is established. However, most studies rely on immunohistological approaches, which have several limitations, such as precluding comprehensive phenotypic analysis. The aim of this study was to perform an alternative analysis of macrophage phenotypes in advanced human atherosclerotic plaques and compare them with their presence in non-atherosclerotic arteries. Atherosclerotic plaques from 70 individuals indicated for carotid endarterectomy, and samples of non-atherosclerotic arterial tissue (renal artery, control group) from 45 living kidney donors were processed to obtain immunocytes and incubated with antibodies (CD45, CD14, CD16, CD36, CD163, and CD206) to be analyzed by flow cytometry. Macrophages in the atherosclerotic plaques tend to express CD16 more intensively than in non-atherosclerotic arterial tissue (transient, CD16 < 0.001, pro-inflammatory, CD16 < 0.001), and the expression is more closely associated with CD36 expression. Both transient and pro-inflammatory macrophages are linked with the CD206CD163 or CD206CD163 phenotype in atherosclerotic plaques, while CD206CD163 dominates within the anti-inflammatory (CD16) population in the control group. Interestingly, when evaluating all macrophages (regardless of CD16 expression), almost all are CD163 in both groups, supporting the critical importance of using a combination of specific markers. Our results provide a deeper insight into macrophage subpopulations in advanced human atherosclerotic plaques compared with those in non-atherosclerotic vessels. Additionally, our data highlight the critical importance of using appropriate techniques, such as flow cytometry, allowing for simultaneous analysis of multiple markers to accurately and comprehensively characterize macrophages within the atherosclerotic plaque.
巨噬细胞极化在动脉粥样硬化形成过程中的重要性已得到证实。然而，大多数研究依赖免疫组织学方法，这种方法存在一些局限性，比如无法进行全面的表型分析。本研究的目的是对晚期人类动脉粥样硬化斑块中的巨噬细胞表型进行另一种分析，并将其与非动脉粥样硬化动脉中的巨噬细胞表型进行比较。对70例因颈动脉内膜切除术而入选的个体的动脉粥样硬化斑块，以及45例活体肾供体的非动脉粥样硬化动脉组织样本（肾动脉，对照组）进行处理，以获取免疫细胞，并与抗体（CD45、CD14、CD16、CD36、CD163和CD206）孵育，然后通过流式细胞术进行分析。动脉粥样硬化斑块中的巨噬细胞比非动脉粥样硬化动脉组织中的巨噬细胞更倾向于强烈表达CD16（瞬时性的，CD16 < 0.001，促炎性的，CD16 < 0.001），并且这种表达与CD36表达的相关性更强。在动脉粥样硬化斑块中，瞬时性和促炎性巨噬细胞都与CD206CD163或CD206CD163表型相关联，而在对照组的抗炎性（CD16）群体中，CD206CD163占主导。有趣的是，当评估所有巨噬细胞（无论CD16表达情况如何）时，两组中几乎所有巨噬细胞都是CD163，这支持了使用特定标志物组合的至关重要性。与非动脉粥样硬化血管中的巨噬细胞亚群相比，我们的结果为晚期人类动脉粥样硬化斑块中的巨噬细胞亚群提供了更深入的见解。此外，我们的数据突出了使用适当技术（如流式细胞术）的至关重要性，这种技术能够同时分析多种标志物，从而准确、全面地表征动脉粥样硬化斑块内的巨噬细胞。
