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- Title
367. Lentivirus Pseudotyped with Envelope Proteins F and HN from Sendai Virus Transduce Airway Epithelium Efficiency and Persistently.
- Authors
Griesenbach, Uta; Mitomo, Katsuyuki; Inoue, Makoto; Somerton, Lucinda; Farley, Raymond; Singh, Charanjit; Tabata, Toshiaki; Ueda, Yasuji; Fujikawa, Satoshi; Washizawa, Kentaro; Hasegawa, Mamoru; Alton, Eric W. F. W.
- Abstract
We have previously shown that recombinant Sendai-virus (SeV) vector transduces airway epithelium efficiently via the apical membrane. This is in part due to cholesterol and sialic acid in the apical membrane which are respectively utilized by the SeV fusion (F) and hemagluttinin-neuraminidase (HN) proteins for receptor- mediated cell attachment, as well as the capacity of SeV to penetrate mucus efficiently. In contrast to SeV, which leads to transient gene expression, integrating lentiviral vectors have the theoretical advantage of persistent expression for the life-time of the cell, and ifstem cell integration should occur, for the lifetime of the animal. We have recently succeeded in generating a F/HN-pseudotyped Simian immunodeficiency virus (SiVagm) vector at high enough titre for in vivo application (>109 transduction units (TU)/ml). A F/HN-SiV- GFP vector (4x108 TU/mouse in 100 ml) was slowly perfused (over 75 min) onto the nasal epithelium of C57Bl/6 mice and transduced airway epithelial cells efficiently, via the apical membrane without the need for chemical pre-conditioning. We choose transduction of the nasal epithelium as proof-of-principle because we aim to extend these studies into CF knockout mice (only the nasal epithelium shows the CF defect in this model). We previously reported detection of GFP expression at 1, 20, 30, 90 days (n=3 mice/group) and 220 days (n=1) after transduction. Here, we further extended these studies to both include higher n numbers and, to identify transduced cells. Importantly, 180 days after transduction 8 out of 10 (6 histological sections 1 mm apart/mouse) mice still expressed GFP. Preliminary experiments on a single animal also showed persistent expression 360 days after transduction, and further animals transduced 270 and 360 days ago, are currently being analysed (n=12–15/group). This duration of gene expression is far longer than the predicted life-span of airway epithelial cells of approximately 100 days. One explanation is that F/HN-SiV vector has transduced progenitor or stem cells in the mouse nose. Morphological analysis showed that GFP positive cells were ciliated airway epithelial cells, and cells being in the olfactory epithelium which based on morphological criteria, appeared to be sensory neuronal cells.Molecular Therapy (2006) 13, S139–S140; doi: 10.1016/j.ymthe.2006.08.426
- Subjects
GENETIC transduction; BACTERIOPHAGES; GENE expression; ISOPENTENOIDS; NEURAMINIDASE; STEM cells
- Publication
Molecular Therapy, 2006, Vol 13, pS139
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1016/j.ymthe.2006.08.426