US EPA

1488783 

Journal Article 

Abstract 

In vivo lentiviral transduction of murine airway epithelium and lung monocytes is enhanced following naphthalene injury 

Aarbiou, J; Van der Wegen, P; Jorna, H; Buijs-Offerman, RM; Spijkers, PP; Scholte, BJ 

2007 

Yes 

Human Gene Therapy
ISSN: 1043-0342
EISSN: 1557-7422 

18 

10 

983 

English 

WOS:000250339500109 

http://://WOS:000250339500109
Exit
 

Efficient transduction of airway epithelium by lentiviral vectors is hampered by innate defense mechanisms in the lung that prevent viral entry. However, treatment with agents that disturb the integrity of lung epithelium has been shown to improve the transduction efficiency of viral vectors. Here we investigated the transduction pattern and efficiency of VSV-G pseudotyped lentiviral vectors after naphthalene-induced Clara cell depletion. Mice were intraperitoneally injected with naphthalene (200 mg/kg), and 2 days later a vector dosage of 1 x 10 super(7) TU of LV-SFFV-GFP or LV-SFFV-LUC in a 50- mu 1 volume was administered intratracheally using a microsprayer device. Oil-injected mice were used as a control. The lentiviral vectors used express either GFP or luciferase under the spleen focus forming virus/murine embryonic stem cell virus promoter/enhancer combination (SFFV). In the naphthalene-LV-SFFV-LUC-treated animals, bioluminescence imaging revealed a lung-restricted transduction pattern and a 7-fold increase in luciferase expression levels when compared to oil-treated controls. Im-munohistochemistry on paraffin sections of naphthalene-LV-SFFV-GFP-treated mice, 1 month after vector administration, showed a 14-fold increase in GFP-positive bronchiolar epithelial cells. GFP-positive epithelial cells are predominantly present in patches of multiple neighboring cells. This suggests that we have selectively transduced a population of epithelial progenitor cells under these conditions. Striking was the abundance of GFP+ lung mononuclear leukocytes (57.5 plus or minus 26.4%) up to 1 month after treatment. The prolonged expression of GFP indicates that a lung-resident hemopoietic stem cell population exists that can be effectively targeted by a lentiviral vector. The methodology used provides promising tools for further study of conducting airways and hemopoietic cells in vivo in lungs using lentiviral vector gene transfer. Supported by Eurocare CF, 6 super(th) framework EEC research program, the Dutch Technology Foundation STW, and the Erasmus "Breedte-Strategie" Program. 

Virology & AIDS Abstracts; Biotechnology and Bioengineering Abstracts; Bioluminescence; Gene transfer; Epithelial cells; Research programs; Embryo cells; Leukocytes (mononuclear); Defense mechanisms; Paraffin; Epithelium; imaging; Promoters; Hemopoiesis; Respiratory tract; Enhancers; Monocytes; Naphthalene; Stem cells; Gene therapy; Injuries; Expression vectors; W 30905:Medical Applications; V 22310:Genetics, Taxonomy & Structure 

15th Annual Congress of the European Society of Gene and Cell Therapy 

Rotterdam, The Netherlands 

October 27-30, 2007