Figure 3.
Mucosal humoral responses. (A) Numbers of OMV-specific IgA-secreting plasma cells in spleens, lungs and blood and (B) numbers of IgA memory cells, that were differentiated into antibody secreting cells (ASC), in spleens and lungs were determined by B-cell ELISpot of 6 mice per group at day 35 and day 56, respectively. Results are indicated as ASC per 5 × 105 cells. Levels of immunoglobulin A (IgA) antibodies directed against Ptx, Prn, FHA, Fim2/3, BrkA, Vag8 and OMV were determined in (C) lungs, (D) nasal wash and (E) serum of 6 mice per group. Results are expressed as fluorescence intensities (FI). (F) Immunoproteomic profiles of pooled (n = 6) pulmonary and nasal IgA were determined using Western blotting using the same scan intensity. In the box plots, the box represent 25th to 75th percentiles, whereas the bars indicate the minimum and maximum value. Horizontal dashed lines represent the limit of detection (background x 3). Significant differences are indicated by *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 obtained using a Mann-Whitney t-test (B-cells) and t-test (antibodies) after log-transformation of data.