In vitro surfactant protein B deficiency inhibits lamellar body formation.

Foster CD, Zhang PX, Gonzales LW, Guttentag SH
Am J Respir Cell Mol Biol. 2003 29 (2): 259-66

PMID: 12649122 · DOI:10.1165/rcmb.2002-0149OC

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.

MeSH Terms (20)

Adenoviridae Blotting, Western Cell Line Cells, Cultured Cell Survival Dose-Response Relationship, Drug Genetic Vectors Humans Lung Microscopy, Electron Models, Genetic Oligonucleotides, Antisense Phenotype Plasmids Pulmonary Surfactant-Associated Protein B Pulmonary Surfactant-Associated Protein C Reverse Transcriptase Polymerase Chain Reaction RNA RNA, Messenger Time Factors

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