Transduction of gammadelta T cells with pseudotyped retroviral vectors

An in-vitro method for transferring one or more nucleic acids sequences comprising one or more transgenes into γδ T cells with a pseudotyped retroviral vector particle or a virus-like particle thereof, comprising the steps:

• a) activating γδ T cells in a sample of leukocytes by adding to said sample an aminobisphosphonate together with one or more cytokine(s) selected from the group consisting of IL-1β, IL-2, IL-7, IL-15, IL-18 and IL-21,
• b) contacting said pseudotyped retroviral vector particle or virus-like particle thereof with said activated γδ T cells, wherein said pseudotyped retroviral vector particle or virus-like particle thereof comprises a modified baboon endogenous retrovirus (BaEV) envelope glycoprotein that binds specifically to and fuses with the surface membrane of hematopoietic cells, thereby genetically modifying said γδ T cells of said sample, and
• c) expanding said genetically modified γδ T cells,
• wherein said pseudotyped retroviral vector particle or virus-like particle thereof comprises at least:
• a chimeric envelope glycoprotein which comprises or consists of a fusion of the transmembrane and extracellular domain of a baboon endogenous retrovirus (BaEV) envelope glycoprotein and the cytoplasmic tail domain of a murine leukemia virus (MLV) envelope glycoprotein;
• or a modified BaEV envelope glycoprotein wherein the cytoplasmic tail domain is devoid of a fusion inhibitory R peptide,
• wherein at least 75% of the γδ T cells obtained following step (c) are CD45RA negative.