The impact of retroviral suicide gene transduction procedures on T cells.
Retroviral vectors encoding the herpes simplex thymidine kinase gene have been used to render T cells sensitive to the prodrug ganciclovir. Such genetically modified T cells have been used in clinical trials for their graft-versus-leukaemia effects following allogeneic haematopoietic stem cell transplantation. In the event of graft-versus-host disease (GVHD) the cells were susceptible to elimination through exposure to ganciclovir. We have investigated the impact of T-cell activation, required for successful retrovirus-mediated gene transfer, on T-cell receptor repertoire profile, subset distribution and antiviral potential. Using a combination of antibodies against CD3 and CD28, T cells were transduced at high efficiency when exposed to retrovirus between 48 and 72 h later. Lymphocytes had undergone up to seven cycles of cell division by the end of the procedure. Although the T-cell receptor Vbeta repertoire was not altered after retroviral transduction, there were notable shifts in subset profiles with an increased proportion of CD45RO cells in transduced populations. T cells continued to proliferate for several days after transduction and were difficult to sustain under the extended culture conditions required to generate virus-specific T cells. These observations may explain the lower than expected levels of GVHD and poor antiviral immunity reported in recent trials.
Qasim
Waseem
W
Molecular Immunology Unit, Institute of Child Health, University College London, London, UK. w.qasim@ich.ucl.ac.uk
King
Douglas
D
Buddle
Jo
J
Verfuerth
Stephanie
S
Kinnon
Christine
C
Thrasher
Adrian J
AJ
Gaspar
Hubert B
HB
D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006086:Graft vs Host Disease; D018380:Hematopoietic Stem Cell Transplantation; D007135:Immunoglobulin Variable Region / Q000032:analysis; D051379:Mice; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D012190:Retroviridae / Q000235:genetics*; D018139:Simplexvirus / Q000201:enzymology*; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation / Q000821:virology*; D013937:Thymidine Kinase / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods
D000998:Antiviral Agents; D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta; D013937:Thymidine Kinase