Lentivirally transduced dendritic cells as a tool for XXXX immunotherapy.

@entity1 @entity5 @entity19 @entity14062 @entity5840
BACKGROUND: Dendritic cells (DC) are the professional antigen-presenting cells of the immune system, fully equipped to prime naive T cells and thus essential components for @entity5 immunotherapy. METHODS: We tested the influence of several elements (cPPT, trip, WPRE, @entity14062 ) on the transduction efficiency of @entity1 DC. @entity1 and @entity19 DC were transduced with tNGFR-encoding lentiviruses to assess the effect of transduction on phenotype and function. @entity1 DC were transduced with lentiviruses encoding huIi80MAGE-A3 and @entity19 DC with huIi80tOVA to test antigen presentation. RESULTS: A @entity14062 ( @entity14062 ) lentiviral vector containing the trip element was most efficient in transducing @entity1 DC. The transduction of DC with trip/ @entity14062 tNGFR encoding lentiviral vectors at MOI 15 resulted in stable gene expression in up to 94.6% ( @entity19 ) and 88.2% ( @entity1 ) of the mature DC, without perturbing viability, phenotype and function. @entity1 huIi80MAGE-A3-transduced DC were able to stimulate @entity5840 -specific CD4(+) and CD8(+) T cell clones and could prime both @entity5840 -specific CD4(+) and CD8(+) T cells in vitro. @entity19 huIi80tOVA-transduced DC were able to present OVA peptides in the context of MHC class I and class II in vitro and induced a strong OVA-specific cytotoxic T lymphocyte response in vivo, that was protective against subsequent challenge with OVA-expressing @entity5 cells. CONCLUSIONS: We show that, using lentiviral vectors, efficient gene transfer in @entity1 and @entity19 DC can be obtained and that these DC can elicit antigen-specific immune responses in vitro and in vivo. The composition of the transfer vector has a major impact on the transduction efficiency.
