Figure 2.

PKCα-Deficient Mice Are Resistant to EAE in a Passive Adoptive-Transfer Model

For adoptive EAE, draining lymph-node cells and splenocytes from MOG_35-55-immunized WT, Prkca^−/−, or PBS-treated WT (control) mice were restimulated with MOG_35-55 in the presence of Th17-cell-polarizing cytokines for 3 days. Data in (A)–(E) were derived from five mice per group. (See also Figure S2). Error bars in (B), (C), (E), and (F) represent ± SEM. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

(A and B) Representative FACS dot plots of IL-17A⁺ and IFN-γ⁺ cells (CD4⁺ gated) are displayed in (A), and the quantification is shown in (B).

(C) IFN-γ (left) and IL-17A (right) levels were measured.

(D and E) Representative FACS dot plots of ROR-γt⁺ cells (CD4⁺ gated) are displayed in (D), and the quantification is shown in (E).

(F) Disease time course of adoptive EAE in WT recipients, reconstituted with Th17-cell-polarized WT (n = 4), Prkca^−/− (n = 4), or control CD4⁺-enriched (n = 2) cells. Of note, control mice injected with cells from PBS-injected WT mice and stimulated in vitro for 3 days with MOG_33-35 under Th17-cell-polarizing conditions did not show any disease signs.

(G) Naive CD4⁺ OT-II T cells were stimulated with OVA_323-339-pulsed DCs under Th17-cell-polarizing conditions. Cells were stained for Vα2TCR and intracellular IL-17A and analyzed by flow cytometry. Representative FACS plots of two independent experiments of three mice per group are shown.