Replication-defective recombinant Semliki Forest virus encoding GM-CSF as a vector system for rapid and facile generation of autologous human tumor cell vaccines.
This paper describes the production of recombinant Semliki Forest virus encoding murine or human granulocyte-macrophage colony-stimulating factor (GM-CSF) and the capacity of these vectors to transduce murine and human tumor cells ex vivo. High-titer stocks (up to 3 x 10(9) particles/ml) of conditionally infective, replication-defective, recombinant SFV particles were generated using the SFV Helper-2 system. It is shown that the recombinant SFV/GM-CSF virus, as well as recombinant SFV carrying the beta-galactosidase reporter gene, efficiently transduce both murine tumor cell lines as well as primary human renal carcinoma cells. Using ELISA's specific for GM-CSF, levels of GM-CSF production by the cells were determined. Levels of murine GM-CSF (mGM-CSF) produced by SFV/mGM-CSF transduced renal cell cancer cultures were equal to or higher than corresponding levels reported in the literature after transduction of similar renal carcinoma cell cultures using a retroviral vector system. The biological activity of GM-CSF was demonstrated by using cells which are dependent on GM-CSF for growth and by using primary bone marrow cells. All the transduced cell cultures (including the human renal cell carcinoma samples) produced GM-CSF for up to at least 4 days after transduction. The results imply that the recombinant SFV system can be used for rapid and facile preparation of autologous cancer cell vaccines.
Withoff
S
S
Department of Medical Microbiology, Molecular Virology Section, University of Groningen and Academic Hospital, Groningen, The Netherlands.
Glazenburg
K L
KL
van Veen
M L
ML
Kraak
M M
MM
Hospers
G A
GA
Störkel
S
S
de Vries
E G
EG
Wilschut
J
J
Daemen
T
T
D000818:Animals; D019677:Artificial Gene Fusion / Q000379:methods; D019496:Cancer Vaccines; D002292:Carcinoma, Renal Cell / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016233:Immunotherapy, Active / Q000379:methods*; D007680:Kidney Neoplasms / Q000276:immunology; D051379:Mice; D012672:Semliki forest virus / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014182:Transplantation, Autologous; D014407:Tumor Cells, Cultured / Q000276:immunology* / Q000378:metabolism; D022701:Virosomes; D001616:beta-Galactosidase / Q000235:genetics
D019496:Cancer Vaccines; D022701:Virosomes; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D001616:beta-Galactosidase