Figure 1.

Re-differentiation and CD4 Modification of T-Lineage Cells from CD4⁺ Th1 Clone-Derived iPSCs Exert HLA Class II-Restricted Responses

(A) Representative flow cytometry profiles of the indicated molecules on the original CD4+ Th clone (SK) and regenerated T cells (iPS-T cells) after 14 days of phytohemagglutinin (PHA)-P stimulation.

(B) Principal component analysis of expression profiles of 146 selected T cell/ILC-related genes (Table S3).

(C) GATA3, RORC, and TBX21 expression in the indicated population. mRNA expression levels were determined by RNA sequencing.

(D) Hierarchical clustering of expressions of 22 selected genes related to ILC subsets.

(E) Cytokine production of the original CD4⁺ Th1 clone (SK) and iPS-T cells. T cells were stimulated with plate-bound control immunoglobulin (immunoglobulin G [IgG]) or anti-CD3 mAb (10 μg/mL) for 24 hr. The indicated cytokines in the culture supernatant were measured by ELISA. Data shown are the means ± SD of triplicate cultures and are representative of two independent experiments.

(F) Representative flow cytometry profiles of CD4 and TCR-Vβ22 expression on the original CD4+ Th1 clone (SK), Mock transduced iPS-T cells (Mock iPS-T cells), and CD4-transduced iPS-T cells (CD4⁺ iPS-T cells).

(G) Proliferative responses of Mock iPS-T cells and CD4⁺ iPS-T cells to antigenic peptide. T cells were co-cultured with autologous PBMCs in the presence of b3a2 peptide (10 μM) and measured by the [³H]thymidine incorporation assay. Data shown are the means and are representative of two independent triplicate experiments.

(H) b3a2 peptide-specific IFN-γ production by Mock iPS-T cells and CD4+ iPS-T cells. Mock iPS-T cells and CD4+ iPS-T cells (1 × 10⁵) were co-cultured for 24 hr with autologous DCs (5 × 10⁴) that had been prepulsed with b3a2 peptide (10 μM). IFN-γ in the culture supernatant (24 hr) was measured by ELISA. Data shown are the means ± SD of triplicate cultures and are representative of two independent experiments.