EZH1/2抑制剂UNC1999对外周血免疫细胞表型的影响

Effects of EZH1/2 Inhibitor UNC1999 on Immunophenotypes of Peripheral Immune Cells

【目的】探讨EZH1/2抑制剂UNC1999对外周血免疫细胞表型的影响。【方法】用CCK-8法检测UNC1999作用于外周血单个核细胞(PBMC)后的细胞存活率;多色流式细胞术分析免疫细胞表型。【结果】相比对照组,UNC1999组经典型单核细胞(CD14^(++)CD16^(−))比例上调[(19.53±1.79)%vs.(66.60±5.02)%,t=13.31,P=0.006],中间型单核细胞(CD14^(++)CD16^(+))和非经典型单核细胞(CD14^(+)CD16^(+))比例下调[(35.08±3.97)%vs.(15.42±2.89)%,t=6.130,P=0.026;(35.50±3.53)%vs.(8.40±3.12)%,t=25.740,P=0.002];CD56^(dim)CD16^(+)、CD56^(dom)CD16^(+)NK细胞亚群比例下调[(3.39±0.86)%vs.(0.27±0.06)%,t=4.882,P=0.040;(80.50±0.64)%vs.(0.63±0.23)%,t=133.100,P<0.0001];初始B细胞比例上调[(10.67±1.76)%vs.(37.99±3.76)%,t=17.690,P=0.003],记忆B细胞、过渡B细胞、浆细胞比例下调[(23.39±4.20)%vs.(11.82±1.90)%,t=7.059,P=0.020;(3.58±0.47)%vs.(1.52±0.56)%,t=26.970,P=0.001;(0.18±0.03)%vs.(0.00±0.00)%,t=8.647,P=0.013];DC比例上调[(0.20±0.05)%vs.(1.38±0.13)%,t=16.500,P=0.004],其中pDC/DC下调[(24.90±1.95)%vs.(12.70±2.11)%,t=7.566,P=0.017],mDC/DC上调[(32.41±13.14)%vs.(60.87±8.43)%,t=8.252,P=0.014];CD8^(+)T细胞亚群中CD8^(+)中枢记忆T细胞、CD8^(+)PD-1^(+)比例上调[(5.62±1.24)%vs.(18.38±2.34)%,t=15.600,P=0.004;(2.50±1.02)%vs.(18.34±2.69)%,t=8.822,P=0.013],CD8^(+)效应记忆T细胞比例下调[(28.27±10.15)%vs.(15.62±9.48)%,t=19.480,P=0.003];CD4^(+)T细胞亚群中CD4^(+)CD27^(+)比例下调[(82.77±2.66)%vs.(56.00±9.01)%,t=5.715,P=0.029]。其余细胞亚群差异无统计学意义(P>0.05)。【结论】UNC1999可以改变PBMC免疫细胞表型。

【Objective】To investigate the effects of EZH1/2 inhibitor UNC1999 on the immune cell phenotypes in peripheral blood of healthy adults.【Methods】CCK8 assay was used to measure the cell viability of peripheral blood mononuclear cells(PBMC).Multicolor flow cytometry was performed to analyze the immunophenotypes.【Results】Compared with DMSO group,UNC1999 group showed increased classical monocytes(CD14^(++)CD16^(−))[(19.53±1.79)%vs.(66.60±5.02)%,t=13.31,P=0.006],decreased intermediate monocytes(CD14^(++)CD16^(+))and non-classical monocytes(CD14^(+)CD16^(+))[(35.08±3.97)%vs.(15.42±2.89)%,t=6.130,P=0.026;(35.50±3.53)%vs.(8.40±3.12)%,t=25.740,P=0.002].The proportions of CD56^(dim)CD16^(+),CD56^(dom)CD16^(+) in UNC1999 group were lower[(3.39±0.86)%vs.(0.27±0.06)%,t=4.882,P=0.040;(80.50±0.64)%vs.(0.63±0.23)%,t=133.100,P<0.0001].UNC1999 group exhibited higher frequency of naive B cells[(10.67±1.76)%vs.(37.99±3.76)%,t=17.690,P=0.003],lower frequency of memory B cells,transitional B cells,plasmablasts B cells[(23.39±4.20)%vs.(11.82±1.90)%,t=7.059,P=0.020;(3.58±0.47)%vs.(1.52±0.56)%,t=26.970,P=0.001;(0.18±0.03)%vs.(0.00±0.00)%,t=8.647,P=0.013].The percentage of DC and mDC/DC was significantly elevated in UNC1999 group[(0.20±0.05)%vs(.1.38±0.13)%,t=16.500,P=0.004;(32.41±13.14)%vs.(60.87±8.43)%,t=8.252,P=0.014],with a significantly decreased percentage of pDC/DC[(24.90±1.95)%vs.(12.70±2.11)%,t=7.566,P=0.017].Higher proportions of CD8^(+) central memory T cells(TCM)and CD8^(+)PD-1^(+)[(5.62±1.24)%vs.(18.38±2.34)%,t=15.600,P=0.004;(2.50±1.02)%vs.(18.34±2.69)%,t=8.822,P=0.013],but lower proportions of CD8^(+) effective memory T cells(TEM)and CD4^(+)CD27^(+) were observed in UNC1999[(28.27±10.15)%vs.(15.62±9.48)%,t=19.480,P=0.003;(82.77±2.66)%vs.(56.00±9.01)%,t=5.715,P=0.029].No statistical difference was found in other cell subsets(P>0.05).【Conclusion】UNC1999 can lead to changes in PBMC immunophenotypes.