Figure 1. PIT induces unresponsiveness in naïve Tg4 cells.

(A, B) B10.PLxC57BL/6 mice received PBS or PIT i.v. 1 day after transfer of naïve CD4⁺ Tg4 cells. EAE was induced 7 days later by immunization with Ac1-9. (A) Mean clinical scores ± SEM. (B) Frequency of CD4⁺ Tg4 cells in the spleen at day 19 post-EAE induction (six mice per group, from one of three experiments giving consistent results). (C) Spleens were sampled four and 7 days after PIT/PBS for analysis of CD4⁺ Tg4 numbers and Foxp3 expression in host and donor CD4⁺ T cells (3–4 mice per group, from one of three experiments giving consistent results). A separate cohort were immunized on day 7 after PIT/PBS and spleens analyzed 10 days later for CD4⁺ Tg4 cell numbers and the production of IFN-γ, IL-17 and IL-10 by splenocytes in response to stimulation with 100 μM Ac1-9 (dotted lines represent cytokine levels for unstimulated controls) (four mice per group, from one of three experiments giving consistent results).

DOI: http://dx.doi.org/10.7554/eLife.03416.003

Figure 1—figure supplement 1. PIT reduces the frequency and number of pro-inflammatory cytokine producing Tg4 cells.

B10.PLxC57BL/6 mice received PBS/PIT 1 day after transfer of naïve CD4⁺Tg4 cells. 7 days later mice were immunized and spleens were taken for analysis 10 days after immunization. (A) Percentage and (B) numbers of IL-17⁺ and IFN-γ⁺ Tg4 donor cells upon recall stimulation with 20 μM Ac1-9. Data are from one of three experiments giving consistent results (four mice per group).

DOI: http://dx.doi.org/10.7554/eLife.03416.004