Figure 3

Siglec-H regulates TLR7-mediated activation of pDCs.

(a–f) The C57BL/6-background WT mice (n = 6), Siglech^kd mice (n = 6) and pDC-ablated mice (n = 6) were injected with or without IMQ and cDCs and pDCs were obtained 24 hrs after injection. (a) The expression of MHC and costimulatory molecules on cDCs and pDCs was analyzed by flow cytometry. Data are MFI ± s.d. from six individual samples in a single experiment. (b–e) CD45.1 OT-II CD4⁺ T cells (b,d) or CD45.1 OT-I CD8⁺ T cells (c,e) were cultured with pDCs (b,c) or cDCs (d,e) in the presence or absence of OVA_323–339 peptide (b,d) or OVA_257–264 peptide (c,e) and the proliferation was measured by [³H]thymidine incorporation. Data are the mean ± s.d. from six individual samples in a single experiment. (f) The expression of CD154 on cDCs and pDCs was analyzed by flow cytometry. Data are MFI ± s.d. from six individual samples in a single experiment. (g,h) CD45.1⁺ OT-II CD4⁺ T cells were cultured with pDCs obtained from WT mice (n = 3) and Siglech^kd mice (n = 3) in the presence or absence of IMQ in combination with OVA_323-339 peptide and various blocking mAbs under T_H1 (g)- or T_H17 (h)-polarized culture conditions for 3 days and intracellular production of IFN-γ (g) or IL-17 (h) in the cultured CD4⁺ T cells was analyzed by flow cytometry. Data are presented as a dot plot and numbers represent the proportion of IFN-γ⁺ cells (g) and IL-17⁺ cells (h) among gated CD4⁺ T cells in each quadrant. *P < 0.01 compared with WT mice. All data are representative of at least three independent experiments.