Table 2.
Relative mRNA expression of TLR (−2, -4, -2/6 and −7) after T-2 toxin treatment in PAM
| negative control | TLR-L | T-2 + TLR-L | |
|---|---|---|---|
| HKLM (TLR2-L) |
1 +/− 0.14 |
76.92 +/− 16.67* |
29.08 +/− 8.16 (*), (t) |
| LPS (TLR4-L) |
1 +/− 0.11 |
17.69 +/− 4.80* |
6.32 +/− 1.46 (*), (t) |
| FSL1 (TLR2/6-L) |
1 +/− 0.22 |
259.90 +/− 75.91* |
81.07 +/− 15.62 (*), (t) |
| Imiquimod (TLR7-L) | 1 +/− 0.20 | 13.66 +/− 3.92* | 7.03 +/− 1.66 (*) |
Primary porcine cells (2 × 106 cells per mL) were exposed or not with 3 nM of T-2 toxin for 1 h before activation during 4 h at 39°C by different TLR-agonists: TLR2-agonist (HKLM); TLR4-agonist (LPS); TLR2/6-agonist (FSL1) and TLR7-agonist (Imiquimod). Relative expression of TLR genes was evaluated by quantitative real time PCR. Data were analyzed by DART-PCR software and gene expression was normalized by the mean of two housekeeping genes (RPL32 and β2-microglobuline). TLR gene expression was reported to unstimulated PAM. After checking for the normal distribution of data carried out with one-way ANOVA test, Student test was conducted to determine the differences among groups. The mean (+/− SEM) of the values obtained with PAM from five different piglets are shown. Significant differences (P < 0.05) between unstimulated cells (negative control) and cells treated by each TLR-L are marked with an asterisk (*). Significant differences (P < 0.05) between TLR-L-activated cells in the presence or not of T-2 toxin treatment are marked with (t).