FIG 3.

AuNP vaccine intranasal immunization elicits robust antigen-specific humoral and cell-mediated responses associated with reduced bacterial adherence. Systemic and mucosa-specific humoral responses were analyzed using sera (A) and intestinal lavage (B) from immunized animals. The intestinal epithelial layer containing intraepithelial lymphocytes was isolated from immunized animals and stimulated in the presence of each antigen for 5 days. Medium-only and anti-CD3/CD28-coated beads served as negative and positive controls, respectively. (C to H) Supernatants from cells stimulated in the presence or absence of antigen were collected and analyzed for cytokine release using a mouse cytokine Bio-Plex. Only the antigens (LpfA1 and LomW) that showed a cytokine response are shown. All humoral and cytokine data are shown as ±SEM from results determined for each group. Significant differences in humoral or cytokine levels were determined via one-way ANOVA followed by Tukey’s post hoc test (*, P < 0.05; **, P < 0.01; ***, P < 0.001).