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

作　　者：刘丹平 池沛冬[2] 梅美华 游莹 黄俊琪 LIU Dan-ping;CHI Pei-dong;MEI Mei-hua;YOU Ying;HUANG Jun-qi(Department of Transplantation,The First Affiliated Hospital//Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology//Guangdong Provincial International Cooperation Base of Science and Technology(Organ Transplantation),Sun Yat-sen University,Guangdong 510080,China;Department of Clinical Laboratory,Sun Yat-Sen University Cancer Center,Guangzhou 510060,China)

机构地区：[1]中山大学附属第一医院器官移植科//广东省器官捐献与移植免疫重点实验室//广东省器官移植国际科技合作基地,广东广州510080 [2]中山大学肿瘤防治中心检验科,广东广州510060

出　　处：《中山大学学报（医学科学版）》2021年第4期494-503,共10页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81871238);广东省器官捐献与移植免疫重点实验室建设项目(2013A061401007,2017B030314018,2020B1212060026);广东省器官移植国际合作基地建设项目(2015B050501002)。

摘　　要：【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.

关 键 词：Zeste增强子同源物1/2 UNC1999 多色流式细胞术 免疫表型 

分 类 号：R392.9[医药卫生—免疫学]