Regulated gene expression in cultured type II cells of adult human lung

Philip L. Ballard, Jae W. Lee, Xiaohui Fang, Cheryl Chapin, Lennell Allen, Mark R. Segal, Horst Fischer, Beate Illek, Linda W. Gonzales, Venkatadri Kolla, Michael A. Matthay

Abstract

Alveolar type II cells have multiple functions, including surfactant production and fluid clearance, which are critical for lung function. Differentiation of type II cells occurs in cultured fetal lung epithelial cells treated with dexamethasone plus cAMP and isobutylmethylxanthine (DCI) and involves increased expression of 388 genes. In this study, type II cells of human adult lung were isolated at ∼95% purity, and gene expression was determined (Affymetrix) before and after culturing 5 days on collagen-coated dishes with or without DCI for the final 3 days. In freshly isolated cells, highly expressed genes included SFTPA/B/C, SCGB1A, IL8, CXCL2, and SFN in addition to ubiquitously expressed genes. Transcript abundance was correlated between fetal and adult cells (r = 0.88), with a subset of 187 genes primarily related to inflammation and immunity that were expressed >10-fold higher in adult cells. During control culture, expression increased for 8.1% of expressed genes and decreased for ∼4% including 118 immune response and 10 surfactant-related genes. DCI treatment promoted lamellar body production and increased expression of ∼3% of probed genes by ≥1.5-fold; 40% of these were also induced in fetal cells. Highly induced genes (≥10-fold) included PGC, ZBTB16, DUOX1, PLUNC, CIT, and CRTAC1. Twenty-five induced genes, including six genes related to surfactant (SFTPA/B/C, PGC, CEBPD, and ADFP), also had decreased expression during control culture and thus are candidates for hormonal regulation in vivo. Our results further define the adult human type II cell molecular phenotype and demonstrate that a subset of genes remains hormone responsive in cultured adult cells.

  • expression profiling
  • dexamethasone
  • cAMP
  • surfactant
  • pepsinogen C
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