Figure 4.

In vitro-induced expression of antimicrobial peptide (AMP) genes in trout B cells by LPS or Aeromonas salmonicida. The IgM⁺ and IgT⁺ B cells were MACS sorted from peripheral blood leukocytes (PBLs) and head kidney leukocytes (HKLs) of trout and incubated with PBS (Ctrl), LPS (25 µg/ml), or heat-killed A. salmonicida (A.sal; cell:bacteria = 1:10) for 8 h at 17°C. Then cells were collected and subjected to total RNA isolation and cDNA synthesis. The relative expression levels of AMP genes in trout IgM⁺ and IgT⁺ B cells under normal and challenged situations were determined by quantitative real-time PCR and normalized against elongation factor 1a (*p < 0.05). Data are representative of three independent experiments (mean ± SD).