Journal: Scientific Reports
Article Title: Bromodomain and extra-terminal protein inhibitors modulate natural killer cell function and differentiation
doi: 10.1038/s41598-025-33437-1
Figure Lengend Snippet: BETi impact function of primary human NK cells. ( A ) Human primary NK cells isolated from healthy donors ( n = 4) unstimulated or stimulated with IL-12 + IL-15 for 24 h with prior (24 h) treatment with BETi (AZD5153 or CPI-203) compared to no drug treatment control. Bar graph of levels of IFNγ secreted into the cell culture supernatant measured by ELISA reported in pg/ml. ** P ≤ 0.01, * P ≤ 0.05, Ratio paired t-test. ( B ) Percentage of live CD14 − CD19 − cells and of total NK cells (live CD14 − CD19 − CD56 + CD16 + ) within the lymphocyte gate (Supplementary Fig. 2) in PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells for a further 5 h and analyzed. ( C ) Representative histogram overlays illustrating expression of granzyme B in NK cells in the indicated conditions, and bar graphs showing the median fluorescence intensity (MFI) of granzyme B staining in NK cells after culture of PBMCs from healthy donors ( n = 6) for 48 h in medium with or without IL-12 + IL-18 in the absence (open circles) or presence of BET inhibitors CPI-203 (black circles) or AZD5153 (blue circles), followed by co-culture for 5 h with CD4.221 cells in the presence of Brefeldin A. ( D – F ) Percentages of CD107a + ( D ), of IFN-γ + ( E ) and of TNF-α + cells ( F ) in total NK cells within PBMCs from healthy donors ( n = 6) incubated in medium only (open circles) or with CPI-203 (black circles) or with AZD5153 (blue circles), in the presence or absence of IL-12 plus IL-18, for 48 h, and then co-cultured with CD4.221 cells in the presence of Brefeldin A for a further 5 h and analyzed. In ( B – F ), each symbol represents data from a single donor; error bars indicate mean ± sem of n = 6 samples. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.00001; one way-ANOVA with Tukey’s post-test.
Article Snippet: Cells were stimulated with 10ng/ml recombinant IL-12 (130-129-720, Miltenyi) plus 100ng/ml recombinant IL-18 (B001-5, R&D Systems) and with 100nM CPI-203 (15479, Cayman Chemical) or 100nM AZD 5153 (15479, Cayman Chemical), or cultured in medium only for 48 h at 37 °C.
Techniques: Isolation, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Incubation, Expressing, Fluorescence, Staining, Co-Culture Assay
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