Figure 5.
Reactivity of the MAGE-specific TCRs against early-passage melanoma cell lines
(A) To determine recognition of early-passage melanoma cell lines (passage ≤10), TCR-T cells were stimulated. All melanomas (MEL) naturally expressed the HLA of interest except for MEL18.07 that was transduced with HLA-A∗01:01. IFN-γ production, as measured by ELISA, was determined after an overnight stimulation in at least two independent experiments. An allo-HLA reactive T cell clone was included for each target HLA as a positive control for HLA expression and stimulatory capacity of target cells. The MAGE-gene expression levels, measured by qPCR, are depicted between brackets as percentage relative to HKGs. Values and error bars represent means and standard deviations of technical duplicates. (B) Cytotoxicity of the TCR-T cells was analyzed by a 6-h 51Cr-release assay in at least two independent experiments. In Figure S5, the positive (allo-HLA reactive clones) and negative (CMV TCR) are shown. Values and error bars represent means and standard deviations of technical triplicates.