FIGURE 3. SV40_IV-TCR transduced T cells recognize antigen in vitro and in vivo.

A, Flow cytometric analysis of SV40_IV-TCR (left panel) or mock (right panel) transduced T cells. The transduction efficiency of the SV40_IV-TCR was determined by calculating the percentage of Vβ9⁺ cells within the population of Vβ-pool⁺ cells (represented in right upper corner of dot plot). Average transduction percentage was 25-40%. B, Functional analysis of SV40_IV-TCR transduced splenocytes. Cells were incubated in the presence of SV40_404-411 (closed circles) or control OVA_257-264 peptide (open circles) at the indicated concentrations. Prior to incubation, a sample of the transduced T cells was stained with K^b-SV40_404-411 tetramers and PE-anti-CD8α antibodies to determine the percentage of SV40_IV-specific CD8⁺ cells. Post peptide stimulation, cells were stained with APC-anti-CD8α, permeabilized and stained with PE-anti-IFNγ. C, Flow cytometric analysis of blood cells of B6 (left panels) and TRAMP mice (right panels) that received 1×10⁵ SV40_IV-TCR transduced T cells (top panels) or no transfer of T cells (bottom panels) followed by an i.n. infection with 1000 p.f.u. of flu-T. Blood was sampled 3-14 days post infection. Circles represent TCR-transduced T cell responses in individual mice; bars indicate averages.