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dc.contributor.authorGilham, David Een
dc.contributor.authorLie-A-Ling, Michaelen
dc.contributor.authorTaylor, Naomien
dc.contributor.authorHawkins, Robert Een
dc.date.accessioned2010-08-10T09:04:36Z
dc.date.available2010-08-10T09:04:36Z
dc.date.issued2010-02
dc.identifier.citationCytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors. 2010, 12 (2):129-36 J Gene Meden
dc.identifier.issn1521-2254
dc.identifier.pmid20033928
dc.identifier.doi10.1002/jgm.1421
dc.identifier.urihttp://hdl.handle.net/10541/109356
dc.description.abstractBACKGROUND: HIV-1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV-1 derived lentiviral vectors for stable gene transfer into mouse T cells. METHODS: Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture. RESULTS: Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV-1 lentiviral vectors also efficiently transduced cytokine-stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully-activated T cells. CONCLUSIONS: The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV-1 lentiviral vectors, opening up prospects for their use in mouse models of gene-modified adoptive cellular therapy.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshCD4-Positive T-Lymphocytes
dc.subject.meshCD8-Positive T-Lymphocytes
dc.subject.meshCell Proliferation
dc.subject.meshCell Separation
dc.subject.meshCytokines
dc.subject.meshGene Expression
dc.subject.meshGenetic Vectors
dc.subject.meshHIV-1
dc.subject.meshHumans
dc.subject.meshLymphocyte Activation
dc.subject.meshMice
dc.subject.meshMice, Inbred BALB C
dc.subject.meshMice, Inbred C57BL
dc.subject.meshPromoter Regions, Genetic
dc.subject.meshSpleen
dc.subject.meshT-Lymphocytes
dc.subject.meshTransduction, Genetic
dc.subject.meshTransgenes
dc.titleCytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors.en
dc.typeArticleen
dc.contributor.departmentCell Therapy Group, Cancer Research UK Department of Medical Oncology, University of Manchester, Manchester Academic Health Science Centre, Manchester, UK. dgilham@picr.man.ac.uken
dc.identifier.journalThe Journal of Gene Medicineen
html.description.abstractBACKGROUND: HIV-1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV-1 derived lentiviral vectors for stable gene transfer into mouse T cells. METHODS: Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture. RESULTS: Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV-1 lentiviral vectors also efficiently transduced cytokine-stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully-activated T cells. CONCLUSIONS: The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV-1 lentiviral vectors, opening up prospects for their use in mouse models of gene-modified adoptive cellular therapy.


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