Published ahead of print on March 20, 2003, doi:10.1165/rcmb.2002-0149OC
© 2003 American Thoracic Society DOI: 10.1165/rcmb.2002-0149OC In Vitro Surfactant Protein B Deficiency Inhibits Lamellar Body FormationDivision of Neonatology, Department of Pediatrics, University of Pennsylvania School of Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania Address correspondence to: Susan Guttentag, M.D., Abramson Research Center 416G, Children's Hospital of Philadelphia, 3516 Civic Center Blvd., Philadelphia, PA 19104-4318. E-mail: guttentag{at}email.chop.edu Surfactant protein (SP) B is essential for normal pulmonary surfactant activity and lamellar body genesis in type 2 cells. However, the role of SP-B in lamellar body genesis is poorly understood. We developed an adenovirus vector expressing antisense SP-B as an alternative in vitro model of SP-B deficiency to begin to explore the role of SP-B in lamellar body genesis. RT-PCR analysis revealed that antisense SP-B expression interfered with translation of endogenous SP-B mRNA. Antisense SP-B expression resulted in reliable in vitro reproduction of many features of SP-B deficiency, including absent mature SP-B and decreased lamellar bodies and SP-C. Light and electron microscopy demonstrated significant reductions in lamellar body number. Western blotting revealed a significant reduction in mature 8-kD SP-B protein and decreased mature SP-C. Our data indicate that antisense SP-B can be effectively used to replicate the SP-B–deficient type 2 cell phenotype in vitro, and provides an attractive alternative to transgenic models for the further study of the role of SP-B in lamellar body genesis.
Abbreviations: enhanced green fluorescent protein, EGFP • glyceraldeyde-3-phosphate deydrogenase, GAPDH • surfactant protein, SP This article has been cited by other articles:
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