Fig. 3.

Quantification of cells secreting TNP-specific IgMs in (A) spleen and (B) kidney 15 and 30 days after the anal administration of the antigens. To determine the amount of cells secreting TNP-specific IgMs, ELISPOT plates were coated with TNP-BSA and developed using a biotinylated anti-trout IgM mAb (clone 4C10). Results are shown as mean number of IgM-secreting cells per 5 × 10⁴ cells +SD (n = 7). Representative wells for each condition are shown. Asterisks denote significantly different values among groups as indicated (*p ≤ 0.05 and ***p ≤ 0.005). (C) Pie charts show the ratio of cells secreting TNP-specific IgM between spleen and kidney at different time points in fish anally immunized with TNP-LPS.