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Low doses of cholera toxin and its mediator cAMP induce CTLA-2 secretion by dendritic cells to enhance regulatory T cell conversion

Fig 4

Both CTlo and cAMP-matured DCs are protective in EAE.

(A) CTlo (0.1 μg/ml) or cAMP (100 μM) treated BM-DCs loaded with 40 μg/ml MOG35-55 peptide were injected i.v. at days -7, -5, -3 (2×106/mouse) before MOG-specific EAE induction at d0. Control mice received PBS injections. Average disease scores were monitored for a total of 10 mice per group from two independent experiments. Error bars represent mean ± SEM. (B), (C) and (D) Statistical analyses of different other clinical parameters derived from A. Error bars represent mean ± SD of pooled results from n = 10 mice. (E) CTlo (0.1 μg/ml), cAMP (100 μM) treated BM-DCs or untreated BM-DCs were injected i.v. at days -7, -5, -3 (2×106/mouse) before MOG-specific EAE induction at d0. Control mice received PBS injections. Average disease scores were monitored for a total of 6 mice per group of two independent experiments. Error bars represent mean ± SEM. (F), (G) and (H) Statistical analyses of different other clinical parameters derived from E. Error bars represent mean ± SD of pooled results from n = 6 mice. (I) Spleen cells of EAE mice treated like in A were analyzed 8 or 16 days after EAE induction for their Foxp3+ Treg relative frequencies or Ki67+ proliferating Foxp3+ Treg frequencies. (J) Like in I, but the spleen cells were re-stimulated with 10 μg/ml MOG35-55 peptide and then the relative frequencies of Treg expansion were determined after 5 days. (K) Like in I, but splenocytes were re-stimulated with graded concentrations of MOG35-55 peptide and after 3 days their supernatants analyzed by ELISA for IL-17A and IFN-γ production. (I-K) Error bars represent mean ± SD of pooled results from n = 6 mice. One Way ANOVA, Dunnett post-test. *p<0.05, **p<0.01, ***p<0.001, ns: not significant.

Fig 4

doi: https://doi.org/10.1371/journal.pone.0178114.g004