Sunday, July 27, 2014 5:38:50 PM
Hum Gene Ther Methods. 2014 Jul 21. [Epub ahead of print]
Targeting both viral and host determinants of HIV entry using a new lentiviral vector co-expressing the T20 fusion inhibitor and a selective CCL5 intrakine.
Petit NY1, Dorgham K, Bault B, Burlion A, Gorochov G, Marodon G.
Author information
1CIMI, INSERM U1135, PARIS, France ; nicolas.petit1984@gmail.com.
Abstract
Numerous strategies targeting early and late steps of the HIV life cycle have been proposed for gene therapy. However, targeting viral and hosts determinants of HIV entry is the only strategy that would prevent viral DNA-mediated CD4 cell death while diminishing the possibility for the virus to escape. To this end, we devised a bicistronic lentiviral vector expressing the membrane bound form of the T20 fusion inhibitor, referred to as the C46 peptide, and a CCR5 super-agonist, modified to sequester CCR5 away from the cell surface, referred to as the P2-CCL5 intrakine. We tested the effects of the vector on HIV infection and replication using the human CEMR5 cell line expressing CD4 and CCR5 and on primary human T cells. Transduced cells expressed the C46 peptide, detected with the 2F5 monoclonal antibody by flow cytometry. Expression of the P2-CCL5 intrakine correlates with lower levels of cell surface CCR5. Complete protection to HIV infection could be observed in cells expressing the protective transgenes. Importantly, we show that the combination of the transgenes was more potent than single transgene alone, showing the interest of expressing two entry inhibitors to inhibit HIV infection. Finally, genetically modified cells possessed a selective advantage over non-modified cells upon HIV challenge in vitro, showing that modified cells were protected from HIV-induced cell death. Our results demonstrate that lentiviral vectors co-expressing the T20 fusion inhibitor and the P2-CCL5 intrakine represent promising tools for HIV gene therapy.
Good luck and GOD bless,
George
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