Lentiviral vectors for T-cell suicide gene therapy: preservation of T-cell effector function after cytokine-mediated transduction.
Retroviral transfer of the Herpes Simplex thymidine kinase (HSVTK) suicide gene to donor T cells has been used as a safety strategy against graft-versus-host disease following allogeneic stem cell transplantation. The feasibility of this strategy in human studies has been demonstrated, but a number of limitations have become apparent. Preactivation of donor lymphocytes using mitogens or monoclonal antibodies is essential for retroviral transduction, but can compromise subsequent T-cell function in vivo. We report the application of lentiviral vectors for transduction of T cells in cytokine culture, without activation through the T-cell receptor. Using vectors encoding either enhanced green fluorescent protein or a truncated CD34/mutant HSVTK fusion selection/suicide construct, we investigated the properties of T cells after gene modification. We found that following cytokine stimulation, a fraction of T cells undergoes division, and transgene expression occurred predominantly in these cells. Antiviral and alloreactive responses were preserved in these populations, and in contrast to fully activated T cells, there was minimal perturbation of regulatory T-cell numbers. We conclude that the use of interleukin-7 for lentiviral transduction offers the greatest potential for gene transfer to T cells without loss of function, and is favored for the clinical production of suicide gene modified T cells.
Qasim
Waseem
W
Molecular Immunology Unit, Institute of Child Health, University College London, London, UK. w.qasim@ich.ucl.ac.uk
Mackey
Taylor
T
Sinclair
Jo
J
Chatziandreou
Ilenia
I
Kinnon
Christine
C
Thrasher
Adrian J
AJ
Gaspar
H Bobby
HB
D016207:Cytokines / Q000494:pharmacology; D005434:Flow Cytometry; D041722:Genes, Transgenic, Suicide / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015851:Interleukin-7 / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D016207:Cytokines; D015851:Interleukin-7; D049452:Green Fluorescent Proteins; D013937:Thymidine Kinase