Figure 2. Detection of potential MiHA specific T-cell populations by pMHC tetramer staining.

These FACS analyses show the detection of MiHA specific T-cell populations through dual-encoding after pMHC tetramer pull down and in vitro expansion. Shown are total CD8⁺ T-cells. All dot plots are shown with bi-exponential axes and display fluorescence intensity for the indicated fluorochromes at the top and right of the plot matrix. Non-pMHC tetramer specific CD8⁺ T-cells are indicated black. Dot plots of pMHC tetramer positive T-cell populations are shown by staining one expanded cell culture with 16 separate panels of up to 25 different dual-color pMHC tetramers. (a) Representative example of pMHC multimer screen panel 5, patient BDY3356. Detection of three dual-labeled potential MiHA specific T-cell populations: P89 ITGAM_RLQ (red), P104 ZFP36L2_RLL (blue) and P109 FMNL1_SLW (green). Frequencies indicate MiHA specific T-cells of total CD8⁺ cells. A selection of 21 potential MiHA specific T-cell populations was made with the highest clinical potential. Selected T-cell populations were derived from allo-SCT patient: OBB1465 (b), JMO2750 (c), BDY3356 (d) and APM4461 (e). Dot plots shown are representative for all detected dual-positive CD8⁺ T-cell populations (red).