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Keratinocyte growth factor induced epithelial proliferation facilitates retroviral-mediated gene transfer to distal lung epithelia in vivo
Journal article   Peer reviewed

Keratinocyte growth factor induced epithelial proliferation facilitates retroviral-mediated gene transfer to distal lung epithelia in vivo

Joseph Zabner, Guoshun Wang, Paul B McCray Jr, Vladimir A Slepushkin, Mordechai Bodner, Helmuth H. G van Es, Patricia Thomas, Doug J Jolly and Beverly L Davidson
The journal of gene medicine, Vol.1(1), pp.22-30
01/1999
DOI: 10.1002/(SICI)1521-2254(199901/02)1:1<22::AID-JGM1>3.0.CO;2-X
PMID: 10738582

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Abstract

Background Cell proliferation, vector titer and accessibility of target cells represent hurdles for efficient gene transfer to lung epithelia in vivo using recombinant murine leukemia (MuLV)-based retroviruses. We tested the hypothesis that the pulmonary epithelium is susceptible to retroviral-mediated gene transfer when stimulated to proliferate by a mitogen, keratinocyte growth factor (KGF). Methods Rats received keratinocyte growth factor (KGF, 2.5 µg/g×4 doses, two consecutive days) intratracheally followed by high titer amphotropic retrovirus expressing β-galactosidase. Gene transfer was assessed five days later. Results KGF stimulated transient proliferation in the bronchiolar and alveolar epithelia (30–40% PCNA positive cells at peak) which decreased to background levels seven days after administration. Gene transfer to epithelia (X-Gal positive cells) occurred more frequently in KGF treated rats, but proliferation exceeded the level of gene transfer. X-gal positive cells were noted in the alveolar epithelium and occasionally in the bronchiolar epithelium. In order to understand the discrepancy between the number of proliferating and transduced cells, primary rat tracheal epithelium cultured at the air-liquid interface was infected from either the apical or basolateral side. Gene transfer was achieved only through basolateral application of vector, suggesting that epithelial polarity represents a barrier to MuLV-based lung gene transfer in vivo. Conclusions KGF transiently stimulates epithelial proliferation in vivo, facilitating MuLV-based gene transfer. Retroviral vectors may encounter multiple barriers which have evolved to defend the lung from infections.
Cystic Fibrosis Pit2 PCNA KGF amphotropic

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