Guselnikova V V, Razenkova V A, Kirik O V, Nikitina I A, Pavlova V S, Zharkina S I, Korzhevskii D E
Institute of Experimental Medicine, St. Petersburg, Russia.
Dokl Biochem Biophys. 2024 Dec;519(1):506-511. doi: 10.1134/S160767292470114X. Epub 2024 Sep 16.
Resident macrophages of different organs have structural and functional features, which can complicate their identification and analysis. A promising candidate for the role of a universal immunohistochemical marker of resident macrophages is the calcium-binding protein Iba-1, a well-known marker of brain microglia. The purpose of this work was to study the possibility of using one variant of antibodies to the Iba-1 protein for the immunohistochemical detection of resident macrophages in the liver, myocardium, lung, and choroid plexus of the rat brain. The study was performed on male Wistar rats (n = 15). It was shown that the use of rabbit monoclonal antibodies against Iba-1 allows highly effective detection of Kupffer cells in the liver, resident macrophages in the myocardium, alveolar and interstitial macrophages in the lung, and Kolmer cells in the choroid plexus of the rat brain. In all cases, the reaction is characterized by a high specificity and the absence of background staining. In contrast to the classical marker of macrophages, the CD68 molecule, the Iba-1 protein is evenly distributed in the cytoplasm of cell bodies and processes. This makes it possible to more fully identify cells using immunostaining for Iba-1, carry out their three-dimensional reconstructions, and study their structural and functional organization. Immunohistochemical reaction against Iba-1 can be successfully used as a universal alternative to other common methods for identifying resident macrophages.
不同器官的常驻巨噬细胞具有结构和功能特征,这可能会使它们的识别和分析变得复杂。钙结合蛋白Iba-1是脑小胶质细胞的一种著名标志物,有望成为常驻巨噬细胞通用免疫组化标志物。本研究的目的是探讨使用一种针对Iba-1蛋白的抗体变体,对大鼠肝脏、心肌、肺和脑脉络丛中的常驻巨噬细胞进行免疫组化检测的可能性。研究选用雄性Wistar大鼠(n = 15)。结果表明,使用抗Iba-1的兔单克隆抗体能够高效检测大鼠肝脏中的库普弗细胞、心肌中的常驻巨噬细胞、肺中的肺泡巨噬细胞和间质巨噬细胞以及脑脉络丛中的科尔默细胞。在所有情况下,反应都具有高度特异性且无背景染色。与巨噬细胞的经典标志物CD68分子不同,Iba-1蛋白均匀分布在细胞体和突起的细胞质中。这使得利用Iba-1免疫染色更全面地识别细胞、进行三维重建以及研究其结构和功能组织成为可能。针对Iba-1的免疫组化反应可成功用作识别常驻巨噬细胞的其他常用方法的通用替代方法。