FIGURE 5.

Truncated sema3A induces regulatory pattern in CD3+T lymphocytes. Primary CD3+T cells were isolated from PBMCs using MACS microbeads and MS column. The cells were cultured in 12-well plates, pre-coated with 10 μg/mL anti-CD3 for 4 h at 37 °C, in addition to 1 μg/mL anti-CD28, for 24 h at 37 °C. After 24 h, 1 μg/mL of P- or T-sema3A or an appropriate amount of a control elution buffer was added for an extra 24 h at 37 °C. On the day of the experiment, cells were stained with fluorescence antibodies and the expression of the following cytokines was evaluated in a NAVIOUS EX flow cytometer. The results were analyzed using Kaluza software, and the statistical significance was calculated using one-way ANOVA and Friedman test (*= p-value<0.05). (A). IL-10 (B). IFN-γ (C). IL-17A.