Fig. 7. CALR-mutant-specific responses in both T_EM and T_CM.

a PBMCs from a healthy individual were enriched for T effector memory cells (T_EM: CD3⁺, CD62L^- CD45RA⁻), T central memory cells (T_CM: CD3⁺, CD62L⁺, CD45RA^-), T_EMRA (CD3⁺, CD62L⁻, CD45RA⁺), and T_naive (CD3⁺, CD62L⁺, CD45RA⁺). Cells were analyzed in an ex vivo IFN-γ ELISPOT for responses against CALRLong4. (Top) Representative wells for each cell fraction show responses to both CALRLong4 and a scrambled negative control peptide; (middle) graph shows the numbers of spot-forming cells in each cell fraction. (Bottom) Analysis of purity of the different T-cell fractions in CD3⁺ gated cells. Due to cell scarcity, T_EM and T_EMRA responses were measured in duplicate and in a single experiment, respectively. T_CM and T_näive responses were measured in triplicate. b PBMCs from healthy individuals were sorted and analyzed as described above. (Top) Representative wells for each cell fraction show responses to both CALRLong4 and a scrambled negative control peptide. (Bottom) Graph shows the number of spot-forming cells in each cell fraction. Due to cell scarcity, we did not perform purity analyses. T_EMRA responses were analyzed in a single experiment, but all other cell fractions were analyzed in triplicate. Error bars display standard error of the mean