In Vivo Induction of Tr1 Cells via Mucosal Dendritic Cells and AHR Signaling
Figure 1
Nasal anti-CD3 induces suppressive Tr1 cells.
A. Tiger mice were nasally treated with IC (clear bars) or anti-CD3 (filled bars) and 72 hrs after the last nasal dose GFP(IL-10) expression by CD4+ T cells in CLN was examined by flow cytometry. This experiment was repeated 4 times with same results. B and C. CD4+CD25-GFP-, CD4+CD25+GFP- or CD4+CD25-GFP+ T cells were sorted from CLN of Tiger mice nasally treated with IC (clear bar) or anti-CD3 (filled bar). Sorted T cells were stimulated in vitro with plate bound anti-CD3 and anti-CD28 antibodies (1 µg/ml each) and IL-10 (B) and (C) IFN-γ were detected in the supernatants by ELISA. Error bars represent standard deviations and P values were calculated by t-test. D. The percentage of CD4+CD25-LAP+ T cells that express IL-10 following nasal anti-CD3 was assessed by intracellular staining. Each symbol represents an individual mouse. E. FACS-sorted Tr1 cells (CD4+CD25-GFP(IL-10)+, clear bar) from CLN of nasal anti-CD3 treated Tiger mice or IL-27 in vitro differentiated Tr1 cells (filled bar) were used in a standard suppression assay with naïve CD4+CD25-GFP- responder T cells at various ratios. To test the role of IL-10 in in vitro suppression, IC or anti-IL-10 (50 µg/ml) neutralizing antibodies were added to co-cultures at 1∶1 ratio.