Characterization and transduction of a retroviral vector encoding human interleukin-4 and herpes simplex virus-thymidine kinase for glioma tumor vaccine therapy.
Vaccination with cytokine-transduced tumor cells represents a potentially important approach to the treatment of central nervous system tumors. We have recently demonstrated the therapeutic efficacy of tumor cell vaccines expressing the murine interleukin 4 (IL-4) and the herpes simplex virus-thymidine kinase in a rat brain tumor model in which nonirradiated vaccine cells can be eliminated by the subsequent administration of ganciclovir. In this report, we demonstrate the construction and characterization of a retroviral vector that encodes human IL-4, neomycin phosphotransferase, and herpes simplex virus-thymidine kinase genes for use in human clinical trials. An MFG-based retroviral vector was used to generate the recombinant retrovirus, TFG-hIL4-Neo-Tk, in which a long terminal repeat-driven polycistronic transcript encodes three cDNAs that are linked and coexpressed using two intervening internal ribosome entry site fragments from the encephalomyocarditis virus. The amphotropic retroviral vector TFG-hIL4-Neo-Tk was then used to infect human primary glioma cultures and skin-derived fibroblasts. After infection and G418 selection, cells produced 89-131 ng/10(6) cells/48 hours of human IL-4, which was determined to be biologically active. Transduced glioma cells were highly sensitive to the cytotoxic effect of ganciclovir. These data demonstrate the suitability of the TFG-hIL4-Neo-Tk vector for therapeutic studies of cytokine-transduced autologous tumor vaccination in patients with malignant gliomas.
Okada
H
H
Department of Neurological Surgery, University of Pittsburgh School of Medicine, Pennsylvania 15213, USA. okadah@msx.upmc.edu
Attanucci
J
J
Tahara
H
H
Pollack
I F
IF
Bozik
M E
ME
Chambers
W H
WH
Lotze
M T
MT
D000818:Animals; D019496:Cancer Vaccines / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D004686:Encephalomyocarditis virus / Q000201:enzymology / Q000235:genetics*; D015774:Ganciclovir / Q000633:toxicity; D005822:Genetic Vectors / Q000627:therapeutic use*; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D005910:Glioma / Q000188:drug therapy / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D006801:Humans; D015847:Interleukin-4 / Q000096:biosynthesis / Q000235:genetics*; D008213:Lymphocyte Activation; D051379:Mice; D010703:Phorbol Esters / Q000494:pharmacology; D051381:Rats; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects
D019496:Cancer Vaccines; D005904:Glial Fibrillary Acidic Protein; D010703:Phorbol Esters; D015847:Interleukin-4; C119222:phorbol-12-myristate; D013937:Thymidine Kinase; D015774:Ganciclovir