Fig. 4

OVA-TCR lentiviruses (LVs) facilitate gene transfer in primary T-cells and provide anti-tumor effects in vitro. a Overview of the process of LVs mediated transduction of murine spleen T-cells with OVA-TCR GFP. Top panel shows the plasmid construct used for transgene expression. Bottom panel shows the overall procedure of T-cell transduction for functional studies. b Overlaid flow cytometry histogram plots and quantification of T-cell transduction depicting OVA-TCR expression in T-cells transduced via LVs at varying multiplicity of infection (MOI). c Quantification of T-cell viability when subjected to transduction at different MOI. d Percentage of B16-OVA tumor cells viability when co-cultured with non-transduced T-cells (UT + B16-OVA) and OVA-TCR expressing transduced T-cells (T + B16-OVA) at 24 h and 48 h at an effector to target ratio of 5:1. e Quantification of IFN-γ and TNF-α released (as assessed by ELISA) in the supernatant of co-culture of T-cells with tumor cells. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, one-way ANOVA with Tukey’s test. OVA-TCR ovalbumin T-cell receptor, GFP green fluorescent protein, EF1α elongation factor 1α, IRES internal ribosome entry sites, eGFP enhanced green fluorescent protein, αCD3 cluster of differentiation 3, αCD28 cluster of differentiation 28, rIL-2 recombinant interleukin-2, IFN-γ interferon-γ, TNF-α tumor necrosis factor-α