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Abstract：
Mass cytometry (MC) is a powerful method for mapping complex cellular systems at single-cell levels, based on the detection of cellular proteins. Numerous studies have been performed using human blood, but there is a lack of protocols describing the processing and labeling of bronchoalveolar lavage fluid (BALF) and nasal polyps (NP) for acquisition by MC. These specimens are essential in the investigation of immune cell characteristics in airway diseases such as asthma and chronic rhinosinusitis with NP (CRSwNP). Here we optimized a workflow for processing, labeling, and acquisition of BALF and NP cells by MC. Among three methods tested for NP digestion, combined enzymatic/mechanical processing yielded maximum cell recovery, viability and labeling patterns compared to the other methods. Treatment with DNAse improved sample acquisition by MC. In a final step, we performed a comparison of blood, BALF and NP cell composition using a 31-marker MC antibody panel, revealing expected differences between the different tissue but also heterogeneity among the BALF and NP samples. We here introduce an optimized workflow for the MC analysis of human NP and BALF, which enables comparative analysis of different samples in larger cohorts. A deeper understanding of immune cell characteristics in these samples may guide future researchers and clinicians to a better disease management.
摘要：
质量细胞术（MC）是在单细胞水平上映射复杂细胞系统的强大方法，基于细胞蛋白质的检测。已经使用人类血液进行了许多研究，但是缺乏描述支气管肺泡灌洗液（BALF）和鼻息肉（NP）的处理和标记的协议，以供MC获取。这些标本对于研究哮喘和慢性鼻窦炎伴NP（CRSwNP）等气道疾病的免疫细胞特征至关重要。在这里，我们优化了处理工作流程，标签,并通过MC获得BALF和NP细胞。在测试NP消化的三种方法中，酶/机械联合处理产生最大的细胞回收率，与其他方法相比，生存力和标记模式。用DNA酶处理改善了MC的样品采集。在最后一步，我们做了血液比较,使用31标记MC抗体组的BALF和NP细胞组成，揭示了不同组织之间的预期差异，以及BALF和NP样品之间的异质性。我们在这里介绍一个优化的工作流程，用于人类NP和BALF的MC分析，这使得在更大的队列中的不同样品的比较分析。更深入地了解这些样本中的免疫细胞特征可能会指导未来的研究人员和临床医生更好地管理疾病。
