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Cytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV‐1 vectors
Author(s) -
Gilham David E.,
LieALing Michael,
Taylor Naomi,
Hawkins Robert E.
Publication year - 2010
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1421
Subject(s) - transduction (biophysics) , biology , viral vector , genetic enhancement , microbiology and biotechnology , transgene , cytokine , t cell , virology , cytotoxic t cell , gene , immunology , immune system , in vitro , genetics , recombinant dna , biochemistry
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.