The interleukin 4 (IL4,) is a cytokine that induces differentiation of naive helper T cells (Th0 cells) to Th2 cells. Upon activation by IL4, Th2 cells subsequently produce additional IL4 in a positive feedback loop. IL4 has many biological roles, including the stimulation of activated B-cell and T-cell proliferation, and the differentiation of B cells into plasma cells. It is a key regulator in humoral and adaptive immunity. IL4 induces B-cell class switching to IgE, and up-regulates MHC class II production. IL4 decreases the production of Th1 cells, macrophages, IFN-gamma, and dendritic cell IL12. The activity of IL4 is usually measured by a cell proliferation assay using TF-1 cells. TF-1 cells were seeded into triplicate wells of 96-well plates at a density of 8,000 cells/well with 2% serum standard 1640 which contains various concentrations of recombinant rat IL4. After incubated for 3 days, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 µl of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 2-4 hours at 37 ℃. Proliferation of TF-1 cells after incubation with IL4 for 3 days observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with recombinant rat IL4 for 3 days. The result was shown in Figure 2. It was obvious that IL4 significantly increased cell viability of TF-1 cells. The ED50 is 13.76 ng/ml.

Figure 2. Cell proliferation of TF-1 cells after stimulated with IL4.