Figure 1
Generation and characterization of allergen-specific T cell stimulator cells and allergen-specific T cell reporter cells.
(a) Scheme illustrating the generation of K562 cell-based engineered APC (left) and expression analysis of surface molecules on these cells by flow cytometry (right). Open histograms: K562 cells; filled histograms: respective engineered APC. (b) Scheme illustrating the generation of Jurkat-based allergen-specific T cell reporter cells (left) and expression analysis of the allergen-specific TCRs using antibodies specific for the Vß-chains of the respective transgenic TCR (right). Open histograms: Staining of allergen-specific reporter cells with isotype control antibody; filled histograms: staining with antibodies specific for the transgenic TCR-Vß chain. (c) Activation of Je6 NF-κB reporters with allergen-specific stimulator cells harboring the appropriate restriction element in absence (open histograms) or presence (grey histograms) of allergenic peptides Art v 123–36 or Bet v 1142–153 (left). T cell reporter specific for Art v 1 and Bet v 1 were cocultured with eAPC expressing the indicated molecules in presence or absence of allergenic peptides. Mean gMFI ± SD of triplicates is shown and experiment is representative for three independently performed experiments. (d) Reporter gene-expression and formation of immunological synapses visualized by cell imaging using chamber slides. K562 stimulator cells expressing HLA-DRB1 (transduced with mCherry for microscopic visualization) are cocultivated with Je6 NF-κB-eGFP-reporter cell line without and with allergenic peptide addition (upper and lower panel, respectively).
