Functional and phenotypic characterization of HA-1 TCR-infusion products. (A) Percentage of cells expressing the CD34 tag within the CD4 (light blue bars) and CD8 (dark blue bars) infusion products. Cell products 1.1, 1.2, and 1.3 are 3 separate products made for patient 1. (B) Degranulation of CD4 (light blue bars) and CD8 (dark blue bars) infusion products when cocultured with primary HA-1⁺ AML (effector:stimulator = 1:1) measured by CD107a flow cytometric assay. Absence of bars indicates that data were not available (eg, product 3, 3.3, 5 CD4). (C-D) Lysis of HA-1 peptide–pulsed (1nM) T2 cells (C) or primary HA-1⁺ AML (D) in 4-hour chromium release assay by CD4 (light blue bars) and CD8 (dark blue bars) cell infusion products (effector:target = 20:1). (E-F) Expression of T-cell differentiation markers (left), activation markers (middle), and activation/inhibition markers (right) in CD4 (n = 11) (E) and CD8 (n = 12) (F) infusion products. Horizontal line represents the median. Each color represents an individual patient. (G) Percentage of CD4 HA-1 TCR-T (in CD4 infusion products, n = 8) or CD8 TCR-T (in CD8 [n = 12] or CD4 infusion products [n = 8]) secreting interferon (IFN)γ (red), IL-2 (blue), tumor necrosis factor (TNF)α (green), individually (unfilled bars) or combinations of cytokines (filled bars) in response to HA-1 peptide–pulsed (1nM) T2 cells by intracellular cytokine staining. Error bars indicate standard error of the mean. Statistics determined by an unpaired t test (2-sided P value: ∗P <. 05, ∗∗P < .01; ∗∗∗P < .001).