Fig. 1

Blocking IL-10 signalling at the time of long HPV16 E7 peptide/MPLA immunization increases the numbers of IL-10 producing cells. Groups of five C57BL/6 mice were immunized as indicated with 50 μg of E7 peptide/15 μg of Monophosphoryl Lipid A (MPLA), 300 μg of anti-IL10R antibodies or control antibodies s.c on days 0 and 14. Splenocytes and draining lymph node cells from immunized mice were harvested and stimulated with 25 ng/ml PMA and 1 μg/ml ionomycin for 6 h in the presence of monensin on 7 days after final immunization. Cells were surface stained for CD3, CD4, and GITR and intracellular stained for IL-10 and IFN-γ. CD3+ cells were gated. Results for cells of draining lymph nodes were shown. A and B: IL-10 secreting CD3 + CD4+ T cells IL-10 (a) FACS plot and (b) summarised data showing IL-10 expression by CD3 + CD4+ T cells. C and D: CD3 + CD4+ T cells secreting IL-10 and IFN-γ dot plots (c) and summarised data from different groups (d) E and F: CD4 + GITR+ T cells secreting IL-10: FACS profile (e) and summarised data from different groups (f). Splenic CD4 + IL-10+ cells were shown in (g)