Cytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors
✍ Scribed by David E. Gilham; Michael Lie-A-Ling; Naomi Taylor; Robert E. Hawkins
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 589 KB
- Volume
- 12
- Category
- Article
- ISSN
- 1099-498X
- DOI
- 10.1002/jgm.1421
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
Background
HIV‐1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV‐1 derived lentiviral vectors for stable gene transfer into mouse T cells.
Methods
Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture.
Results
Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV‐1 lentiviral vectors also efficiently transduced cytokine‐stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully‐activated T cells.
Conclusions
The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV‐1 lentiviral vectors, opening up prospects for their use in mouse models of gene‐modified adoptive cellular therapy. Copyright © 2009 John Wiley & Sons, Ltd.