Functional long-term thymidine kinase suicide gene expression in human T cells using a herpesvirus saimiri vector.
Herpesvirus saimiri transforms human T lymphocytes to stable growth and persists episomally without genomic integration and without virus production. The transformed T cells retain essential features of their parental cells including the MHC-restricted antigen specificity which may be useful for applications in adoptive immunotherapy. In order to improve the biological safety of such vectors, the prodrug activating gene thymidine kinase of herpes simplex virus was inserted into the genome of herpesvirus saimiri by homologous recombination. After infection with wild-type or cloned recombinant viruses, T cells from tamarin monkeys and from humans were transformed to stable growth. Thymidine kinase-expressing transformed T cells were efficiently eliminated in the presence of low concentrations of ganciclovir. This elimination mechanism remained fully functional over an observation period of 12 months. The potentially immunogenic neomycin resistance gene expression cassette was deleted from the genome of established mutant viruses by using the prokaryotic Cre/LoxP recombination system. At any time during the course of a therapeutic application, thymidine kinase-expressing transformed human T cells might be eliminated after administration of ganciclovir. In principle, this function could be useful for the T cell-dependent immunotherapy of resistant blood cancer while avoiding the risk of uncontrolled graft-versus-host disease.
Hiller
C
C
Institut für Klinische und Molekulare Virologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
Wittmann
S
S
Slavin
S
S
Fickenscher
H
H
D000212:Acyclovir / Q000627:therapeutic use; D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000627:therapeutic use; D000998:Antiviral Agents / Q000627:therapeutic use; D002478:Cells, Cultured; D005472:Fluorouracil / Q000627:therapeutic use; D015774:Ganciclovir / Q000627:therapeutic use; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics*; D006801:Humans; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012452:Saguinus; D013601:T-Lymphocytes / Q000201:enzymology*; D013937:Thymidine Kinase / Q000235:genetics*; D019076:Transgenes
D000964:Antimetabolites, Antineoplastic; D000998:Antiviral Agents; D013937:Thymidine Kinase; D015774:Ganciclovir; D005472:Fluorouracil; D000212:Acyclovir