Production and characterization of a bicistronic Moloney-based retroviral vector expressing human interleukin 2 and herpes simplex virus thymidine kinase for gene therapy of cancer

M. Pizzato, E. Franchin, P. Calvi, R. Boschetto, M. Colombo, S. Ferrini, G. Palu

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Gene-based therapeutic strategies for cancer mainly include augmentation of immunotherapeutic and chemotherapeutic approaches. In this study we report the design and functional assay of a novel bicistronic Moloney-based retroviral vector expressing human interleukin-2 (IL-2) and herpesvirus thymidine kinase (tk) through a cap-dependent translation and an internal ribosome entry site (IRES)-regulated translation, respectively. This construct has the potential for allowing combination of cytokine and suicide gene therapy, especially in areas such as the brain, composed of post-mitotic cells refractory to transduction by type C retroviral vectors. Accordingly, human glioma cells were used as targets for gene transfer after selecting a packaging cell clone that produced a reasonable titer of recombinant virus and expressed high levels of IL-2 and tk transcripts. Although transduction efficiency was reduced in glioma cells as compared with murine NIH 3T3 cells, transgene expression was effectively achieved. Transduced glioma cells were sensitive to ganciclovir and secreted around 1000 U/ml IL-2 in the culture supernatants. Simultaneous production of IL-2 and tk in vivo by genetically treated tumor cells would hopefully potentiate the effect of gangiclovir-induced metabolic suicide, possibly by boosting the immune response associated with tumor debulking or by amplifying the bystander response.

Original languageEnglish
Pages (from-to)1003-1007
Number of pages5
JournalGene Therapy
Volume5
Issue number7
Publication statusPublished - 1998

Fingerprint

Thymidine Kinase
Simplexvirus
Genetic Therapy
Interleukin-2
Glioma
Neoplasms
Suicide
NIH 3T3 Cells
Ganciclovir
Herpesviridae
Product Packaging
Viral Load
Transgenes
Genes
Clone Cells
Cytokines
Brain

Keywords

  • Gene therapy
  • Interleukin-2
  • IRES
  • Retroviral vector
  • Thymidine kinase

ASJC Scopus subject areas

  • Genetics

Cite this

Production and characterization of a bicistronic Moloney-based retroviral vector expressing human interleukin 2 and herpes simplex virus thymidine kinase for gene therapy of cancer. / Pizzato, M.; Franchin, E.; Calvi, P.; Boschetto, R.; Colombo, M.; Ferrini, S.; Palu, G.

In: Gene Therapy, Vol. 5, No. 7, 1998, p. 1003-1007.

Research output: Contribution to journalArticle

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AU - Colombo, M.

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