Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection

D. Sangiolo, M. Lesnikova, R. A. Nash, M. C. Jensen, A. Niktine, H. P. Kiem, G. E. Georges

Research output: Contribution to journalArticle

Abstract

Several clinical studies of gene-modified T cells have shown limited in vivo function of the cells, immunogenicity of the transgene, and lack of a selective advantage for genemodified T cells. To address these problems, we developed a lentiviral vector (LV) that provides a selectable, proliferative advantage and potentially decreases immunogenicity for transduced T cells. The bicistronic vector expressed two genes linked with an internal ribosomal entry site. One gene is a variant of the inosine monophosphate dehydrogenase 2, inosine monophosphate dehydrogenase (IMPDHIY), conferring resistance to the immunosuppressive drug mycophenolate mofetil (MMF). The other is a suicide gene, herpes simplex virus thymidine kinase (HSV-TK), rendering proliferating cells sensitive to ablation with ganciclovir, fused to the selectable transmembrane marker DCD34 (DCD34/TK). Cells transduced with V-DCD34/TK.IMPDHIY were efficiently enriched by immunomagnetic selection for CD34, proliferated in 0.5-5 μM MMF, and were killed by 0.5 25 μg ml1 ganciclovir. We demonstrate efficient selection and killing of gene-modified cells and suggest LV-ΔCD34/TK.IMPDHIY- transduced T cells could be used to facilitate allogeneic hematopoietic cell engraftment. The expression of IMPDHIY would allow in vivo selection with MMF, and DCD34/TK expression would allow rapid and safe elimination of transduced T cells if raft-versus-host disease developed.

Original languageEnglish
Pages (from-to)1549-1554
Number of pages6
JournalGene Therapy
Volume14
Issue number21
DOIs
Publication statusPublished - Nov 2007

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Mycophenolic Acid
Inosine Monophosphate
Ganciclovir
Oxidoreductases
T-Lymphocytes
Genes
Thymidine Kinase
Simplexvirus
Immunosuppressive Agents
Transgenes
Suicide

ASJC Scopus subject areas

  • Genetics

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Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection. / Sangiolo, D.; Lesnikova, M.; Nash, R. A.; Jensen, M. C.; Niktine, A.; Kiem, H. P.; Georges, G. E.

In: Gene Therapy, Vol. 14, No. 21, 11.2007, p. 1549-1554.

Research output: Contribution to journalArticle

Sangiolo, D. ; Lesnikova, M. ; Nash, R. A. ; Jensen, M. C. ; Niktine, A. ; Kiem, H. P. ; Georges, G. E. / Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection. In: Gene Therapy. 2007 ; Vol. 14, No. 21. pp. 1549-1554.
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