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Viruses 2010, 2(8), 1577-1588; doi:10.3390/v2081577
Article

A VSV-G Pseudotyped Last Generation Lentiviral Vector Mediates High Level and Persistent Gene Transfer in Models of Airway Epithelium In Vitro and In Vivo

1,†
, 1,‡
, 1,2
 and 1,2,*
1 Institute for the Experimental Treatment of Cystic Fibrosis, H.S. Raffaele, via Olgettina 58, 20132 Milan, Italy 2 Department of Biomedical Sciences, University of Foggia, 71122 Foggia, Italy Present address: Laboratorio Clinical Trials, Istituto di Ricerche Farmacologiche “Mario Negri”, 20156 Milan, Italy. Present address: ReSearch Pharmaceutical Services, Inc., Fort Washington, PA 19034, USA.
* Author to whom correspondence should be addressed.
Received: 21 June 2010 / Revised: 29 July 2010 / Accepted: 29 July 2010 / Published: 2 August 2010
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Abstract

The aim of this work was to evaluate the efficiency and duration of gene expression mediated by a VSV-G pseudotyped last generation lentiviral (LV) vector. We studied LV efficiency in ex-vivo models of respiratory epithelial cells, obtained from bronchial biopsies and nasal polyps, by GFP epifluorescence and cytofluorimetry. In vivo efficiency and persistence of gene expression was investigated by GFP immunohistochemistry and luciferase activity in lung cryosections and homogenates, respectively, upon intranasal and intratracheal administration protocols in C57Bl/6 mice. Both primary bronchial and nasal epithelial cells were transduced up to 70-80% 72 hr after the LV infection. In vivo nasal luciferase expression was increased by lysophosphatidylcholine pre-treatment of the nose. Conversely, the bronchial epithelium was transduced in the absence of any pre-conditioning treatment and luciferase expression lasted for at least 6 months without any decline. We conclude that a last generation LV vector is a promising gene transfer agent in the target organ of genetic and acquired lung diseases, as in the case of cystic fibrosis.
Keywords: lentivirus; cystic fibrosis; GFP; luciferase; lysophosphatidylcholine; airway epithelial cells; lung; mice lentivirus; cystic fibrosis; GFP; luciferase; lysophosphatidylcholine; airway epithelial cells; lung; mice
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Copreni, E.; Palmieri, L.; Castellani, S.; Conese, M. A VSV-G Pseudotyped Last Generation Lentiviral Vector Mediates High Level and Persistent Gene Transfer in Models of Airway Epithelium In Vitro and In Vivo. Viruses 2010, 2, 1577-1588.

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