Respiratory and Sleep Medicine, Royal Children's Hospital, Melbourne, Victoria, Australia.
Respiratory Diseases.
Am J Respir Cell Mol Biol. 2020 Aug;63(2):152-159. doi: 10.1165/rcmb.2019-0453MA.
Childhood pulmonary diseases not only cause childhood morbidity and mortality but also can cause long-term pulmonary impairment. The clinical management of many childhood pulmonary diseases is hampered by a limited understanding of the underlying pathophysiological mechanisms. Flow cytometry, which can be used to phenotype individual cell populations or isolate cells for downstream analysis, represents a crucial technology that can help to elucidate the pathophysiology of these conditions. Here, we describe a flow cytometry-based method for purification and characterization of cell populations in BAL from children. This includes assessment of the effect of cryopreservation on cell phenotype and frequency, a knowledge gap recently identified by an American Thoracic Society report on flow cytometry in lung samples. To our knowledge, this is the first study to simultaneously quantify alveolar macrophages, T cells (CD4 and CD8), B cells, natural killer cells, dendritic cells, granulocytes, and monocytes (CD16/CD16) in the BAL of children. The protocols described can be used to advance investigation of the pathophysiology of childhood pulmonary diseases.
儿童肺部疾病不仅会导致儿童发病率和死亡率增加，还可能导致长期肺部损伤。由于对许多儿童肺部疾病的潜在病理生理机制了解有限，其临床管理受到了阻碍。流式细胞术可用于表型鉴定单个细胞群体或分离细胞进行下游分析，是一种重要的技术，可以帮助阐明这些疾病的病理生理学。在这里，我们描述了一种基于流式细胞术的方法，用于从儿童的 BAL 中纯化和鉴定细胞群体。这包括评估冷冻保存对细胞表型和频率的影响，这是美国胸科学会（American Thoracic Society）关于肺部样本流式细胞术的报告中最近确定的一个知识空白。据我们所知，这是第一项同时定量儿童 BAL 中肺泡巨噬细胞、T 细胞（CD4 和 CD8）、B 细胞、自然杀伤细胞、树突状细胞、粒细胞和单核细胞（CD16/CD16）的研究。所描述的方案可用于深入研究儿童肺部疾病的病理生理学。
