Figure 4.

Functional Analysis of T-iPS-DCs

(A–C) Production of (A) TNF-α, (B) IL-6, and (C) IL-12p70 in mature T-iPS-DCs and mo-DCs stimulated for 72 hr with TNF-α, lipopolysaccharide, and IL-4 or OK432 (10 μg/mL, 25 μg/mL) or CD40L (100 ng/mL, 1 μg/mL). Values are means ± SD. ^∗p < 0.05 by Mann-Whitney test.

(D) Flow cytometric analysis of CD11c gated T-iPS-DCs incubated with green-labeled zymosan for 60 min at 37°C (solid line) or 4°C (dotted line).

(E) Index mean value of the ratio of zymosan incorporation at 37°C and 4°C. Values are means ± SD. N.S., not significant.

(F) T-iPS-DCs or mo-DCs were co-cultured with allogeneic T cells for 5 days. Proliferation of T cells was measured based on bromodeoxyuridine uptake in the last 20 hr of the culture. ^∗p < 0.05 by Mann-Whitney's test. Data were run in triplicate.

(G) T-iPS-DCs or mo-DC pre-incubated with or without M3R peptide (10 μg/mL) were co-cultured with the M3R-reactive T cell line (5 × 10⁴ cells/well). Autoreactive CD4+ T cell responses were evaluated. ^∗p < 0.05 by Mann-Whitney test. N = 3 independent experiments.

(H) T-iPS-DCs or mo-DCs (5 × 10³ cells/well) were co-cultured with allogeneic T cells (3 × 10⁴ cells/well) of healthy donors (HC1-3) or patients with SS (SS1-3) for 5 days. Proliferation of T cells was measured.

(I) T-iPS-DCs (1 × 10⁴ cells/well) were pre-incubated with or without M3R peptide (10 μg/mL) and co-cultured with auto-CD4+ T cells (1 × 10⁵ cells/well). M3R-reactive CD4+ T cells were evaluated.

Error bars denote SD.