Figure 5: The mucin-like peptide Fhmuc induces specific immune response characterized by a strong production of IFNγ. A) C57BL/6 mice were immunized i.p. with Fhmuc (10 g) or PBS (control group) at days 0, 14, 28. At day 42, spleens and PECs were removed and cultured with Fhmuc peptide (10 g/ml) or FhTE (20 g/ml). Secreted cytokines (IFNγ, IL-5) levels were tested on culture supernatants by interleukin- specific sandwich ELISA assays. B) Mice were inoculated i.p. Fhmuc (10 g/ml) and 2 h later LPS (1 g/ml) was administrated. IL12/23p40 and IFNγ levels were tested in sera after 6 h of LPS administration. C) BMDCs were incubated with Fhmuc (10 g/ml) and 2 h later LPS was added and cells were incubated overnight at 37°C. Then IL-12/23p40, IL-6 and IL-10 levels were detected on culture supernatants. D) BMDCs treated with Fhmuc and LPS overnight at 37°C were washed and inoculated i.p. at 1×106 cells/mouse. Ten days later, spleens and PECs were removed and cultured in the presence of Fhmuc (10 g/ml) or FhTE (50 g/ml). IFNγ or IL-5 was detected on culture supernatants by specific ELISA. Results are expressed as mean value of triplicates (± SD, indicated by error bars) and are representative of two different experiments. Asterisks represent differences statistically significant (*p< 0.01, **p< 0.05).