Figure 2.

CD40L^high Population in iPS-T Cells Shows High Responsiveness to TCR Stimulation

(A and B) CD40L expression of indicated iPS-T cells on day 13 after PHA-P stimulation. Mock iPS-T cells or CD4⁺ iPS-T cells were stimulated with PHA-P and cultured in the absence (A) or presence (B) of IL-2. The frequency of CD40L-positive cells is shown in the upper right corner of each panel.

(C and D) Expression of CD40L, CD4, and TCR-Vβ22 on the subpopulations are shown. CD40L high and low populations under IL-2 (100 U/mL) and IL-15 (5 ng/mL) were separated from Mock iPS-T cells (C) and CD4⁺ iPS-T cells (D) by flow cytometry sorting and expanded by PHA-P stimulation.

(E and F) Surface CD40L expression on different subpopulations (E; CD40L high and F; CD40L low) stimulated with plate-bound control IgG or anti-CD3 mAb (10 μg/mL). The original CD4⁺ Th1 clone (SK) served as a control. CD40L (red) and isotype-matched controls (gray) are shown.

(G) Cytokine production by the indicated population stimulated with plate-bound control IgG or anti-CD3 mAb (10 μg/mL). The original CD4⁺ Th1 clone (SK) served as control. (H) Cytokine production by CD40L^high CD4⁺ iPS-T cells (1 × 10⁵) co-cultured with THP1 cells (5 × 10⁴) expressing HLA-DR9 and BCR-ABL p210 gene. (G and H) The indicated cytokines in the culture supernatant (24 h) were measured by a bead-based multiplex immunoassay. Data shown are the means ± SD of triplicate cultures and are representative of two independent triplicate experiments.