Ishikawa Shingo, Miyazawa Masataka, Zibiki Yoshinori, Kamikakimoto Rie, Hobo Seiji
Division of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University, Osaka, Japan.
Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan.
J Vet Med Sci. 2022 Apr 15;84(4):548-557. doi: 10.1292/jvms.21-0522. Epub 2022 Feb 10.
Understanding the immune dynamics in the respiratory mucosa of calves is necessary for a good management of bovine respiratory disease. Immune dynamics in the respiratory mucosa in humans and experimental animals has been assessed by flow cytometric analysis of bronchoalveolar lavage fluid (BALF); however, few reports have addressed this subject in calves. The aim of this study was to establish a universal method to analyze bronchoalveolar lavage fluid (BALF) by flow cytometry and to obtain basic knowledge of bovine respiratory mucosal immune dynamics. We investigated the immune cell populations in BALF and evaluated the surface antigen expression of alveolar macrophages in calves using flow cytometer. To further analyze the surface antigen variation observed in alveolar macrophages in detail, stimulation assays were performed in vitro. BALF cells were separated into three distinct populations based on their light scatter plot, which were considered to be macrophages, lymphocytes, and neutrophils. In most individuals, most of the BALF immune cells were alveolar macrophages, but an increased proportion of lymphocytes and neutrophils was observed in some individuals. Analysis of each surface antigen expression in alveolar macrophages showed that CD21 and MHC class II expression changed in response to changes in the leukocyte population. Moreover, when alveolar macrophages were stimulated with interferon-γ in vitro, the expression of CD21 was drastically reduced and MHC class II was increased, suggesting that functional changes in alveolar macrophages themselves are involved in the immune dynamics.
了解犊牛呼吸道黏膜中的免疫动态对于良好管理牛呼吸道疾病至关重要。人和实验动物呼吸道黏膜中的免疫动态已通过支气管肺泡灌洗液（BALF）的流式细胞术分析进行评估；然而，关于犊牛这一主题的报道很少。本研究的目的是建立一种通过流式细胞术分析支气管肺泡灌洗液（BALF）的通用方法，并获得牛呼吸道黏膜免疫动态的基础知识。我们使用流式细胞仪研究了犊牛BALF中的免疫细胞群体，并评估了肺泡巨噬细胞的表面抗原表达。为了进一步详细分析肺泡巨噬细胞中观察到的表面抗原变化，进行了体外刺激试验。根据光散射图，BALF细胞被分为三个不同的群体，分别被认为是巨噬细胞、淋巴细胞和中性粒细胞。在大多数个体中，大多数BALF免疫细胞是肺泡巨噬细胞，但在一些个体中观察到淋巴细胞和中性粒细胞的比例增加。对肺泡巨噬细胞中每种表面抗原表达的分析表明，CD21和MHC II类分子的表达随着白细胞群体的变化而改变。此外，当肺泡巨噬细胞在体外受到干扰素-γ刺激时，CD21的表达急剧降低，而MHC II类分子的表达增加，这表明肺泡巨噬细胞自身的功能变化参与了免疫动态。
