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.

小儿肺部疾病不仅会导致小儿发病率和死亡率，而且还会引起长期肺损伤。对许多儿童肺部疾病的临床管理受到对潜在病理生理机制的有限理解的阻碍。流式细胞术可用于对单个细胞群进行表型分析或分离细胞以进行下游分析，它是一项关键技术，可帮助阐明这些疾病的病理生理。在这里，我们描述了一种基于流式细胞仪的方法，用于纯化和表征儿童BAL中的细胞群体。这包括评估冷冻保存对细胞表型和频率的影响，这是美国胸科协会最近关于流式细胞术在肺样品中的报告所发现的知识缺口。据我们所知，⁺ / CD16 ⁻）在儿童的BAL中。所描述的协议可用于推进对儿童肺部疾病的病理生理学研究。