Flow cytometric analysis of intracellular cytokines in unstimulated PBMCs in the presence of CD99HIgG.

PBMCs were cultured in the presence or absence of CD99HIgG or HIgG control. T cells, monocytes, B cells and NK cells were gated for the determination of intracellular IL-6, TNF-α and IFN-γ production. The gating strategies were showed in S2 Fig. (A) The graphs indicate fold change of geometric mean fluorescence intensity (GeoMFI) of cytokine positive cells in the indicated conditions (n = 6 for T cells, n = 3 for monocytes, B cells and NK cells). (B) The graphs indicate the subtraction of percentage of cytokine positive cells in the indicated conditions (n = 6 for T cells, n = 3 for monocytes, B cells and NK cells). 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, **p ≤ 0. 01, ***p ≤ 0. 001, ns = not statistically significant.