8 research outputs found
Adhesion of Subsets of Human Blood Mononuclear Cells to Endothelial Cells In Vitro, as Quantified by Flow Cytometry
Intercellular adhesion molecule-1 on cultured human epithelial cell lines: influence of proinflammatory cytokines
Effect of glucocorticosteroids on tumour necrosis factor-alpha-induced intercellular adhesion molecule-1 expression in cultured primary human nasal epithelial cells
Soluble Intercellular Adhesion Molecule-1 and E-selectin in Patients with Asthma Exacerbation
Taming the neutrophil: calcium clearance and influx mechanisms as novel targets for pharmacological control
Novel flow cytometry approach to identify bronchial epithelial cells from healthy human airways
Sampling various compartments within the lower airways to examine human bronchial epithelial cells (HBEC) is essential for understanding numerous lung diseases. Conventional methods to identify HBEC in bronchoalveolar lavage (BAL) and wash (BW) have throughput limitations in terms of efficiency and ensuring adequate cell numbers for quantification. Flow cytometry can provide high-throughput quantification of cell number and function in BAL and BW samples, while requiring low cell numbers. To date, a flow cytometric method to identify HBEC recovered from lower human airway samples is unavailable. In this study we present a flow cytometric method identifying HBEC as CD45 negative, EpCAM/pan-cytokeratin (pan-CK) double-positive population after excluding debris, doublets and dead cells from the analysis. For validation, the HBEC panel was applied to primary HBEC resulting in 98.6% of live cells. In healthy volunteers, HBEC recovered from BAL (2.3% of live cells), BW (32.5%) and bronchial brushing samples (88.9%) correlated significantly (p = 0.0001) with the manual microscopy counts with an overall Pearson correlation of 0.96 across the three sample types. We therefore have developed, validated, and applied a flow cytometric method that will be useful to interrogate the role of the respiratory epithelium in multiple lung diseases