Figure 7: IL-27 does not change pathogenicity of OVA-reactive Tc17 cells.

Flow cytometric analysis of transgenic TCR expression on CD8⁺ T cells from C57BL/6 and OT-I mice (Vα2⁺Vβ5.1⁺ TCR is marker of OT-I T cells) (A). Splenic OT-I Tc17 cells were initially differentiated with TGF-β+IL-6 and OVA_257–264 peptide for 3 days and then reactivated in the presence of IL-23 ± IL-27. CD8⁺ T cells from culture were enriched with magnetic beads and injected i.p. into RIP-mOVA C57BL/6 mice. A portion of Tc17 cells used for the injection was stimulated with PMA and ionomycin in the presence of Golgiplug for 4 h, stained and analyzed by flow cytometry for IL-17A and IFN-γ (B; upper panel). Blood glucose levels >200 mg/dL (dotted line on plot) were considered a sign of hyperglycemia (C). Five days after the transfer, RIP-mOVA C57BL/6 mice were sacrificed and pancreas-draining LN cells or splenocytes were stimulated with PMA and ionomycin in the presence of Golgiplug for 4 h, then stained and analyzed by flow cytometry for IL-17A and IFN-γ on gated OT-I cells identified by Vα2/Vβ5.1 TCR expression, as shown in A (B; medium and lower panels). Data are representative of two experiments with at least 6 mice per group (error bars, s.e.m; ns = not significant).