Flow cytometric analysis of intracellular cytokines produced by monocytes, B cells and NK cells upon OKT3 stimulated PBMCs in the presence of CD99HIgG.

PBMCs were stimulated with anti-CD3 mAb (OKT3) in the presence or absence of CD99HIgG or HIgG control. Cells were gated for the determination of intracellular IL-6, TNF-α and IFN-γ in monocytes, B cells and NK cells. The gating strategies for monocytes, B cells and NK cells were shown in S2 Fig. (A) The graphs indicate fold change of geometric mean fluorescence intensity (GeoMFI) (compared to no added protein condition) of cytokine positive cells in the indicated conditions (n = 4). (B) The graphs indicate the subtraction of percentage of cytokine positive cells (% positive cells in tested–no protein) in the indicated conditions (n = 4). Each dot represents each tested subject and the horizontal lines connect between each subject. Statistical analysis was performed by unpaired t test. *p ≤ 0.05; ns = not statistically significant.