IL-2 (Interleukin-2) is a cytokine produced by T-cells in response to antigenic or mitogenic stimulation. IL-2 is a type of signaling molecule in the immune system, that is required for both T-cell and B-cell proliferation and other activities crucial to regulation of the immune response.Therefore, in order to detect the bioactivity of recombinant pig IL-2, spleen single suspensions were prepared, activated with conA (final concentration 3 ug/ml). Cells were collected after 72h and washed with hanks.Then mouse splenic lymphocytes were were seeded into triplicate wells of 96-well plates at a density of 10,000 cells/well with or without the addition of various concentrations of recombinant pig IL-2. After incubated for 96h,?cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). 10 μl of CCK-8 solution was added to each well of the plate, the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37 °C. Proliferation of Splenic lymphocytes cells after incubation with IL-2 for 96h observed by inverted microscope was shown in Figure 1. The dose-effect curve of recombinant pig IL-2 was shown in Figure 2. It was obvious that recombinant pig IL-2 significantly promoted cell proliferation of Splenic lymphocytes cells. The EC50 for this effect is typically 0.0072-0.0079 ug/ml.

Figure 2. The dose-effect curve of IL-2 on Splenic lymphocytes cells