title: Targeting both viral and host determinants of human immunodeficiency virus entry, using a new lentiviral vector coexpressing the T20 fusion inhibitor and a selective CCL5 intrakine.
abstract: Numerous strategies targeting early and late steps of the HIV life cycle have been proposed for gene therapy. However, targeting viral and host 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 superagonist, 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 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 against HIV infection could be observed in cells expressing the protective transgenes. Importantly, we show that the combination of the transgenes was more potent than either transgene alone, showing the interest of expressing two entry inhibitors to inhibit HIV infection. Last, genetically modified cells possessed a selective advantage over nonmodified cells on HIV challenge in vitro, showing that modified cells were protected from HIV-induced cell death. Our results demonstrate that lentiviral vectors coexpressing the T20 fusion inhibitor and the P2-CCL5 intrakine represent promising tools for HIV gene therapy.
authors.affiliation: 1 Sorbonne Universités , UPMC Univ Paris 06, CR7, Centre d'Immunologie et des Maladies Infectieuses (CIMI), F-75013, Paris, France .
authors.forename: Nicolas
authors.initials: N
authors.lastname: Petit
authors.forename: Karim
authors.initials: K
authors.lastname: Dorgham
authors.forename: Béatrice
authors.initials: B
authors.lastname: Levacher
authors.forename: Aude
authors.initials: A
authors.lastname: Burlion
authors.forename: Guy
authors.initials: G
authors.lastname: Gorochov
authors.forename: Gilles
authors.initials: G
authors.lastname: Marodon
mesh_terms: D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D015658:HIV Infections / Q000628:therapy; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000821:virology; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: C515020:C46 HIV-1 fusion inhibitory peptide; D018946:Chemokine CCL5; D011993:Recombinant Fusion Proteins