Determination of cytokine-producing splenocytes.

<p>Cytokines (IFN-γ and IL-4) were enumerated by ELISPOT at 7 days after the final immunization from the splenocytes. (A and C) Number of spots specific to IFN-γ per 5×10⁵ splenocytes were stimulated with sM2 proteins (left) and M2 peptides (right) of the intranasally (top) and orally (bottom) immunized groups. (B and D) Number of spots specific to IL-4 per 5×10⁵ splenocytes stimulated with sM2 protein (left) and M2 peptide (right) of the intranasally (top) and orally (bottom) immunized groups. NC denotes negative controls (non-immunized), and PC denotes positive controls, the cells stimulated with mitogen. Bars denote mean ± S.D. The asterisk indicates a significant difference between pgsA-CTA1-sM2/<i>L. casei</i> and other groups (*<i>P</i><0.05).