Figure 5.

HLA-G dimer inhibits activation of CD8⁺ T cells and GZMB expression in vitro. A) Representative flow cytometry plots from HVs, showing gating strategy used to identify CD25⁺-expressing activated CD8⁺ T cells for indicated groups (n = 5/group). Histograms depict GZMB expression of CD8⁺ CD25⁺ cells (red line). Filled histogram shows the isotype control (Iso). The flow cytometry dot plots shown are from 1 of 2 separate experiments. Graphical summary shows the frequency (%) of CD8⁺ CD25⁺ GZMB⁺ cells per group. B) Immunofluorescence microscopy showing unstimulated and stimulated PBMCs from HVs with or without HLA-G dimer pretreatment (n = 5/group). Number of GZMB-positive cells was determined by immunofluorescence staining with anti-GZMB mAb conjugated with FITC; nuclear staining is shown by DAPI (Original magnification, ×20. Scale bars, 50 μm). Graphical summary shows the number of GZMB-positive cells counted per group. Data are representative of 2 separate experiments. N.s., not significant. Data are shown as means ± sd. *P < 0.05, ***P < 0.001.