Fig. 3.

Comparison of MHC class I tetramer staining in situ and MHC class I tetramer staining of cell suspensions or cytotoxic activity of cultured T cells of same DTH biopsies from melanoma patients vaccinated with peptide-loaded DC. Cryosections stained with MHC class I tetramers (red), anti-CD8 (green) and DAPI (blue). DTH biopsy (DC loaded with gp100:280) from patient 1. a Positive HLA-A2.1-gp100₂₈₀ tetramer staining (red). b HLA-A2.1-gp100₁₅₄ tetramer staining. c Chromium release assay showing the lysis of HLA-A2.1-positive BLM cells, transfected with gp100 and not the control transfectant G250 by T cells cultured from the DTH biopsy. DTH biopsy (DC loaded with 3 peptides and KLH) from patient 3. d Positive HLA-A2.1-gp100₂₈₀ tetramer staining. e Cryosection from the same area as shown in d stained with HLA-A2.1-tyrosinase tetramer. f Tetramer analysis by flow cytometry of DTH-derived T cells. Depicted is the side scatter on the x-axis (double staining with CD8 FITC might interfere with the tetramer staining) and on the y-axis MHC class I tetramer PE staining. DTH biopsy (DC loaded with 3 peptides and KLH) from patient 4. g Positive HLA-A2.1-gp100₁₅₄ tetramer staining. h Cryosection from the same area as shown in g stained with HLA-A2.1-gp100₂₈₀ tetramer. i Chromium release assay showing the lysis of HLA-A2.1-positive T2 cells, loaded with gp100:154 and not T2 cells loaded with irrelevant, gp100:280 or tyrosinase peptides by T cells cultured from the DTH biopsy. j Tetramer analysis by flow cytometry of DTH-derived T cells. Depicted is the forward scatter on the x-axis (double staining with CD8 FITC might interfere with the tetramer staining) and on the y-axis MHC class I tetramer PE staining