Department of Small Animal Clinical Sciences (Aulakh) and Department of Veterinary Biomedical Sciences (Maltare, Phuong Khanh, Singh), University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B4.
Can J Vet Res. 2021 Jul;85(3):161-169.
The cell surface protein CD34 is expressed in various human tissues and cells, including hematopoietic stem cells, vascular endothelial cells, mucosal dendritic cells, mast cells, eosinophils, microglia, fibrocytes, muscle satellite cells, and platelets. There is a lack of data on the expression of CD34 in canine and porcine tissues. Therefore, we designed a series of immunoblotting, immunohistochemistry, and immunofluorescence experiments to observe CD34 expression in murine, canine, and porcine lungs. We used a rabbit antibody (clone EP373Y) to target the conserved human CD34 C-terminal region and validated its immunoreactivity against mouse lung homogenates. The data showed diffuse bronchiolar and alveolar epithelial localization of CD34 protein in normal murine, canine, and porcine lungs. At 9 or 24 h after bacterial endotoxin exposure, murine CD34 protein shifted to specific bronchoalveolar cells with a punctate pattern, as quantified by CD34 fluorescence. Specific porcine bronchoalveolar cells and leukocytes had significant CD34-positive immunostaining after H3N1 influenza infection. Thus, our study provides fundamental data on the expression of CD34 in lungs and validates an antibody for use in further experiments in these animal species.
细胞表面蛋白 CD34 表达于多种人类组织和细胞中,包括造血干细胞、血管内皮细胞、黏膜树突状细胞、肥大细胞、嗜酸性粒细胞、小胶质细胞、成纤维细胞、肌肉卫星细胞和血小板。关于 CD34 在犬科和猪科组织中的表达数据较少。因此,我们设计了一系列免疫印迹、免疫组织化学和免疫荧光实验,以观察 CD34 在鼠、犬和猪肺中的表达。我们使用针对人 CD34 C 端保守区的兔抗体(克隆 EP373Y),并验证其对鼠肺匀浆的免疫反应性。数据显示,在正常鼠、犬和猪肺中,CD34 蛋白弥漫性定位于细支气管和肺泡上皮。在细菌内毒素暴露 9 或 24 小时后,通过 CD34 荧光定量,鼠 CD34 蛋白转移到具有点状模式的特定细支气管肺泡细胞。在感染 H3N1 流感后,猪的特定细支气管肺泡细胞和白细胞有明显的 CD34 阳性免疫染色。因此,我们的研究提供了 CD34 在肺部表达的基本数据,并验证了一种抗体可用于这些动物物种的进一步实验。