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. 2020 May 5;217(7):e20191369. doi: 10.1084/jem.20191369

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© 2020 Santos et al.

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Figure S2. — Post-trial assessment of MA-specific CD8 T cells from two HLA-A2⁺ patients 1.5–2 yr after DC vaccination. PBMC samples from two patients were used to determine the frequency of circulating dextramer-specific CD8 T cells as well as to examine checkpoint expression in CD8 and MA-specific CD8 T cells for patient 10 and patient 18 (A), 1.5 and 2 yr after DC vaccination, respectively. Frequencies shown for each dextramer were calculated by subtracting the negative control dextramer (Neg dex) frequency for each sample tested. Raw data histograms are shown with isotype controls for coexpression of other proteins as labeled. (B) Measurement of IgG4 in serum-free supernatants of HD DC transduced with Ad5.hPD1Ab. Two different HD monocyte-derived DC preparations were cultured for 5 d, harvested, and transduced with 500 or 1,000 MOI Ad5.hDP1Ab for 3 h before replating using serum-free AIMV media. Supernatant aliquots were taken each day as indicated after DC transduction. The concentration of human IgG4 in supernatants was quantified using a human IgG4 ELISA kit (Invitrogen) per manufacturer’s instructions. (C) Graphs show distribution of PD-1 and CTLA-4 protein expression in circulating lymphocytes grouped according to clinical response. The Wilcoxon rank sum test was used for calculating P values. *, P ≤ 0.05; **, P ≤ 0.01. ns, not significant; Tyros, tyrosinase.