Figure 4.
IDLV expressing mTRP2 as a self-antigen induces low specific T cells. (a) Expression of mTRP2 in cell lines. Mock Lenti-X cells, pTY2CMVmTRP2W-transfected Lenti-X cells and B16F10 cells were fixed and stained with anti-TRP2 antibody (green) and DAPI (blue) and analyzed by CLSM. Images represent a 3D reconstruction from 14 single optical sections. Results from one representative experiment are shown for each analysis. Scale bars are shown for each figure. (b) Pilot study of immunogenicity. C57Bl6 mice (n = 3) were immunized once with 10 × 106 RT units/mouse of either IDLV-mTRP2 or LV-mTRP2. The TRP2-specific T cell-mediated immune response was evaluated by IFNγ-ELISpot assay at two and four weeks in blood. Blood cells were stimulated with H-2Kb restricted TRP2 9mer peptide (SVYDFFVWL). Data are expressed as specific spot forming cells (SFC) per million cells. Box plots show mean ± SEM and single values from each immunized mouse.