Immunology and Allergy Group (GC01), Maimonides Biomedical Research Institute of Cordoba (IMIBIC), University of Cordoba, Reina Sofia University Hospital, Avda Menedez Pidal s/n, 14004 Cordoba, Spain.
Cardiovascular Pathology Group (GA09), Maimonides Biomedical Research Institute, Reina Sofia University Hospital, University of Cordoba, Avda Menedez Pidal s/n, 14004 Cordoba, Spain.
Int J Mol Sci. 2023 Jan 22;24(3):2194. doi: 10.3390/ijms24032194.
Aortic stenosis (AS) is a frequent cardiac disease in old individuals, characterized by valvular calcification, fibrosis, and inflammation. Recent studies suggest that AS is an active inflammatory atherosclerotic-like process. Particularly, it has been suggested that several immune cell types, present in the valve infiltrate, contribute to its degeneration and to the progression toward stenosis. Furthermore, the infiltrating T cell subpopulations mainly consist of oligoclonal expansions, probably specific for persistent antigens. Thus, the characterization of the cells implicated in the aortic valve calcification and the analysis of the antigens to which those cells respond to is of utmost importance to develop new therapies alternative to the replacement of the valve itself. However, calcified aortic valves have been only studied so far by histological and immunohistochemical methods, unable to render an in-depth phenotypical and functional cell profiling. Here we present, for the first time, a simple and efficient cytometry-based protocol that allows the identification and quantification of infiltrating inflammatory leukocytes in aortic valve explants. Our cytometry protocol saves time and facilitates the simultaneous analysis of numerous surface and intracellular cell markers and may well be also applied to the study of other cardiac diseases with an inflammatory component.
主动脉瓣狭窄（AS）是一种常见的老年人心血管疾病，其特征为瓣膜钙化、纤维化和炎症。最近的研究表明，AS 是一种活跃的炎症性动脉粥样硬化样过程。特别是，已经有人提出，存在于瓣膜浸润中的几种免疫细胞类型，有助于其退化和向狭窄进展。此外，浸润的 T 细胞亚群主要由寡克隆扩增组成，可能针对持续存在的抗原特异性。因此，对参与主动脉瓣钙化的细胞进行特征分析，并分析这些细胞对哪些抗原作出反应，对于开发替代瓣膜置换的新疗法至关重要。然而，迄今为止，仅通过组织学和免疫组织化学方法研究了钙化的主动脉瓣，这些方法无法对表型和功能细胞进行深入分析。在这里，我们首次提出了一种简单有效的基于细胞术的方案，该方案可用于鉴定和定量分析主动脉瓣标本中的浸润性炎症性白细胞。我们的细胞术方案节省了时间，并便于同时分析许多表面和细胞内细胞标志物，并且很可能也适用于研究具有炎症成分的其他心脏疾病。
