Human Airway Epithelial Cell Determinants of Survival and Functional Phenotype for Primary Human Mast Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 102; no. 40; pp. 14380 - 14385
• Main Authors: Hsieh, F. H., Sharma, P., Gibbons, A., Goggans, T., Erzurum, S. C., Haque, S. J., Austen, K. Frank
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 04-10-2005; National Acad Sciences
• Subjects: Allergies; Analysis of Variance; Asthma; Biological Sciences; Cell Culture Techniques; Cell Differentiation - immunology; Cell Differentiation - physiology; Cell Survival - immunology; Cell Survival - physiology; Cells; Chymases; Coculture techniques; Connective tissues; Cysteine - metabolism; Cytokines; Cytokines - metabolism; DNA Primers; Eicosanoids; Enzyme-Linked Immunosorbent Assay; Epithelial cells; Genotype & phenotype; Histamine release; Humans; Immunohistochemistry; Leukotrienes - metabolism; Lungs; Mast cells; Mast Cells - metabolism; Mast Cells - physiology; Phenotype; Phenotypes; Respiratory Mucosa - metabolism; Respiratory Mucosa - physiology; Respiratory system; Reverse Transcriptase Polymerase Chain Reaction; Serine Endopeptidases - metabolism; Stem Cell Factor - metabolism; Time Factors; Tryptases
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0503948102

Mast cells (MCs) are found in increased numbers at airway mucosal surfaces in asthmatic patients. Because human airway epithelial cells (HAECs) actively participate in airway inflammatory responses and are in direct contact with MCs in the mucosa, we hypothesized that HAEC-MC interactions may contribute to the differentiation and survival of MCs in the airway mucosa. Here, we show that HAECs express mRNA and protein for soluble and membrane-bound stem cell factor, releasing soluble stem cell factor into the cell culture supernatant at a concentration of 5.9 ± 0.1 ng per 106HAEC. HAECs were able to support MC survival in coculture in the absence of any exogenous cytokines for at least 4 d. Before the initiation of coculture, MCs were uniformly tryptase and chymase ( MCTC) double positive, but by 2 d of coculture the majority of MCs expressed tryptase ( MCT) alone. MCs supported in coculture generated low amounts of cysteinyl-leukotrienes (cys-LT) after FcεRI-dependent activation (0.2 ± 0.1 ng of cys-LT per 106cells) and required priming with IL-4 and IL-3 during coculture to achieve a quantity of cys-LT generation within the range expected for human lung mucosal MC (26.5 ± 16 ng of cys-LT per 106cells). In these culture conditions, HAECs were able to direct mucosal MC protease phenotype, but T cell-derived Th2 cytokines were required for the expression of a functional airway MC eicosanoid phenotype. Thus, distinct cell types may direct unique aspects of reactive mucosal MC phenotype in the airways.