Flow cytometric analysis of bronchoalveolar lavage fluid immune dynamics in calves

The Journal of Veterinary Medical Science

article

Shingo ISHIKAWA¹  Masataka MIYAZAWA²  Yoshinori ZIBIKI²  Rie KAMIKAKIMOTO²  Seiji HOBO² 

[1]Division of Veterinary Science, Graduate School of Life and Environmental Biosciences, Osaka Prefecture University

[2]Joint Faculty of Veterinary Medicine, Kagoshima University

关键词: alveolar macrophage;    bronchoalveolar lavage;    calf;    flow cytometry;    mucosal immunology;   

DOI  :  10.1292/jvms.21-0522

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

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.

【 授权许可】

Unknown