IFN-γ production was initially inhibited, followed by pan-inhibition of Th1/Th2/Th17 cytokines production in splenocytes of BALB/c mice intraperitoneally infected with <i>F</i>. <i>pedrosoi</i> sclerotic cells.

<p>(A and B) BD Cytometric Beads Array (CBA) murine cytokine kit was introduced to measure the Th1/Th2/Th17 cytokine levels in culture supernatants of splenocytes from BALB/c mice at 10 days, 30 days and 50 days after intraperitoneally inoculation with <i>F</i>. <i>pedrosoi</i>-spores, -hyphae or transformed sclerotic cells respectively (n = 12 per group). The BALB/c mice of the same age intraperitoneally inoculated with normal saline were set as inoculation control (n = 12). Before measurement, the splenocytes from BALB/c mice were adjusted to 2×10⁶ cells in a volume of 2 mL RPMI1640 plus 10% FCS, and were pretreated with 1×Cell Stimulation Solution at 37°C for 6h. (A) Seven bead populations with distinct APC-A fluorescence intensities had been coated with capture Abs specific for IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17A and IL-10 proteins, as indicated by A1-A7 in turns. The concentration of specific cytokine mentioned above can be revealed by the fluorescence intensity of PE-conjugated detection Ab, and be calculated according to the standard curve established by cytokine standards using FCAP Array software. The assay diluent incubated with beads and PE-conjugated Ab was set as blank control. (B) Graph showing the concentration (pg/mL) of Th1/Th2/Th17 cytokines produced by splenocytes from uninfected and <i>F</i>. <i>pedrosoi</i>-infected BALB/c mice mentioned above at the indicated days post-inoculation. Data represent the mean±SEM (n = 4 per group at each indicated time point) and statistical analysis was performed using Univariate ANOVA and LSD-t test. Significant: ^* P<0.05; Highly Significant: ^** P<0.01.