目的研究细胞程序性死亡配体-1(programmed cell death protein ligand-1,PD-L1)功能抑制调控免疫活化影响ApoE^(-/-)小鼠动脉粥样硬化发生发展的机制。方法将24只ApoE^(-/-)小鼠随机分为正常组,高脂组和高脂^(+)抗PD-L1单抗组,通过高...目的研究细胞程序性死亡配体-1(programmed cell death protein ligand-1,PD-L1)功能抑制调控免疫活化影响ApoE^(-/-)小鼠动脉粥样硬化发生发展的机制。方法将24只ApoE^(-/-)小鼠随机分为正常组,高脂组和高脂^(+)抗PD-L1单抗组,通过高胆固醇饲料喂养建立动脉粥样硬化(atherosclerosis)模型。实验动物饲养70 d后,分离各组实验动物血管(主动脉根部至腹主动脉)及肝脏组织,进行油红O染色;HE染色检测肝组织病理改变;ELISA检测血清中总胆固醇(CHO)、甘油三酯(TG)、高密度脂蛋白(HDL-c)、低密度脂蛋白(LDL-c)和炎症因子(IFN-γ、TNF-α、IL-1β)含量。流式细胞计数检测肝脏淋巴细胞(CD4^(+)、CD8^(+)、CD4^(+)IFN-γ^(+)和CD8^(+)IFN-γ^(+)T细胞)。RT-PCR检测肝脏组织IFN-γ、TNF-α、IL-1β、CD4和CD8表达。结果与高脂组比较,给予抗PD-L1单抗后促血管壁及肝脏脂质累积并上调血清及肝组织CHO、TG、LDL-c和HDL-c含量。高脂饲养条件下给予抗PD-L1单抗促血清和肝组织谷丙转氨酶(GPT)和谷草转氨酶(GOT)含量升高,但是对碱性磷酸酶(AKP)含量没有影响。高脂饲养条件下给予抗PD-L1单抗促血清和肝脏组织IFN-γ、TNF-α和IL-1β含量升高。高脂饲养条件下给予抗PD-L1单抗抑制CD4表达及促CD8表达。高脂饲养条件下给予抗PD-L1单抗促肝脏CD8^(+)T和CD8^(+)IFN-γ^(+)T细胞活化,但是对CD4^(+)IFN-γ^(+)T细胞活化没有影响。结论高脂饲养条件下给予抗PD-L1单抗通过活化肝脏CD8^(+)IFN-γ^(+)T细胞损伤肝脏功能加重动脉粥样硬化。展开更多
Objective:The expression of programmed death 1(PD-1)on CD8^(+)T cells is associated with their activation and exhaustion,while CD57 serves as a senescence marker.The impact of PD-1^(+)and CD57^(+)CD8^(+)T cells on the...Objective:The expression of programmed death 1(PD-1)on CD8^(+)T cells is associated with their activation and exhaustion,while CD57 serves as a senescence marker.The impact of PD-1^(+)and CD57^(+)CD8^(+)T cells on the prognosis of patients with advanced high-grade serous ovarian cancer(HGSOC)remain unclear.Methods:We assessed the percentages of PD-1^(+)and CD57^(+)CD8^(+)T cells in tumor-infiltrating lymphocytes(TILs,n=85)and tumor ascites lymphocytes(TALs,n=87)using flow cytometry.The optimal cutoffs for these markers in TILs and TALs were determined through the log-rank maximization method.Gene expression analysis elucidated the tumor immune microenvironment(TIME,n=36).Results:Patients with higher PD-1^(+)CD8^(+)TILs(>87.8%)exhibited longer platinum-free interval(PFI)and overall survival(OS).In contrast,those with elevated CD57^(+)CD8^(+)TALs(>28.69%)were more likely to experience chemotherapy and had lower complete remission rates,shorter PFI and OS.PD-1^(+)CD8^(+)TILs are primarily displayed an effector memory state with strong proliferative and secretory capabilities.Approximately 50%of CD57^(+)CD8^(+)TALs were terminally differentiated,exhibiting significantly impaired proliferation.Based on the proportions of PD-1^(+)CD8^(+)TILs and CD57^(+)CD8^(+)TALs,patients were categorized into good,median and poor prognosis groups,with median PFI of 47.78,27.29 and 11.96 months,respectively(P<0.0001).Median OS for these groups was not reach,49.23 and 30.92 months,respectively(P<0.0001).Patients with poor prognosis exhibit significantly reduced CD8^(+)T cell proportion and increased M2 macrophage in the TIME,alongside downregulation of multiple T cell activation-related pathways.Conclusions:Lower levels of PD-1^(+)CD8^(+)TILs and higher CD57^(+)CD8^(+)TALs,assessed prior to treatment,correlated with poor prognosis and suppressive TIME in advanced HGSOC.展开更多
文摘目的研究细胞程序性死亡配体-1(programmed cell death protein ligand-1,PD-L1)功能抑制调控免疫活化影响ApoE^(-/-)小鼠动脉粥样硬化发生发展的机制。方法将24只ApoE^(-/-)小鼠随机分为正常组,高脂组和高脂^(+)抗PD-L1单抗组,通过高胆固醇饲料喂养建立动脉粥样硬化(atherosclerosis)模型。实验动物饲养70 d后,分离各组实验动物血管(主动脉根部至腹主动脉)及肝脏组织,进行油红O染色;HE染色检测肝组织病理改变;ELISA检测血清中总胆固醇(CHO)、甘油三酯(TG)、高密度脂蛋白(HDL-c)、低密度脂蛋白(LDL-c)和炎症因子(IFN-γ、TNF-α、IL-1β)含量。流式细胞计数检测肝脏淋巴细胞(CD4^(+)、CD8^(+)、CD4^(+)IFN-γ^(+)和CD8^(+)IFN-γ^(+)T细胞)。RT-PCR检测肝脏组织IFN-γ、TNF-α、IL-1β、CD4和CD8表达。结果与高脂组比较,给予抗PD-L1单抗后促血管壁及肝脏脂质累积并上调血清及肝组织CHO、TG、LDL-c和HDL-c含量。高脂饲养条件下给予抗PD-L1单抗促血清和肝组织谷丙转氨酶(GPT)和谷草转氨酶(GOT)含量升高,但是对碱性磷酸酶(AKP)含量没有影响。高脂饲养条件下给予抗PD-L1单抗促血清和肝脏组织IFN-γ、TNF-α和IL-1β含量升高。高脂饲养条件下给予抗PD-L1单抗抑制CD4表达及促CD8表达。高脂饲养条件下给予抗PD-L1单抗促肝脏CD8^(+)T和CD8^(+)IFN-γ^(+)T细胞活化,但是对CD4^(+)IFN-γ^(+)T细胞活化没有影响。结论高脂饲养条件下给予抗PD-L1单抗通过活化肝脏CD8^(+)IFN-γ^(+)T细胞损伤肝脏功能加重动脉粥样硬化。
基金supported by National Natural Science Foundation of China(No.82372888)National Key Research and Development Program of China(No.2022YFC2704000)+6 种基金National Natural Science Foundation of China(No.82273383)the Capital’s Funds for Health Improvement and Research(No.2020-2-4098)Youth program of Beijing Municipal Natural Science Foundation(No.7204328)Clinical Medicine Plus X-Young Scholars Project,Peking University(No.PKU2022LCXQ020)Key Clinical Project of Peking University Third Hospital(No.BYSY2022050)Key Clinical Projects of Peking University Third Hospital(No.BYSYZD2021006)Key Clinical Projects of Peking University Third Hospital(No.BYSYZD2019034).
文摘Objective:The expression of programmed death 1(PD-1)on CD8^(+)T cells is associated with their activation and exhaustion,while CD57 serves as a senescence marker.The impact of PD-1^(+)and CD57^(+)CD8^(+)T cells on the prognosis of patients with advanced high-grade serous ovarian cancer(HGSOC)remain unclear.Methods:We assessed the percentages of PD-1^(+)and CD57^(+)CD8^(+)T cells in tumor-infiltrating lymphocytes(TILs,n=85)and tumor ascites lymphocytes(TALs,n=87)using flow cytometry.The optimal cutoffs for these markers in TILs and TALs were determined through the log-rank maximization method.Gene expression analysis elucidated the tumor immune microenvironment(TIME,n=36).Results:Patients with higher PD-1^(+)CD8^(+)TILs(>87.8%)exhibited longer platinum-free interval(PFI)and overall survival(OS).In contrast,those with elevated CD57^(+)CD8^(+)TALs(>28.69%)were more likely to experience chemotherapy and had lower complete remission rates,shorter PFI and OS.PD-1^(+)CD8^(+)TILs are primarily displayed an effector memory state with strong proliferative and secretory capabilities.Approximately 50%of CD57^(+)CD8^(+)TALs were terminally differentiated,exhibiting significantly impaired proliferation.Based on the proportions of PD-1^(+)CD8^(+)TILs and CD57^(+)CD8^(+)TALs,patients were categorized into good,median and poor prognosis groups,with median PFI of 47.78,27.29 and 11.96 months,respectively(P<0.0001).Median OS for these groups was not reach,49.23 and 30.92 months,respectively(P<0.0001).Patients with poor prognosis exhibit significantly reduced CD8^(+)T cell proportion and increased M2 macrophage in the TIME,alongside downregulation of multiple T cell activation-related pathways.Conclusions:Lower levels of PD-1^(+)CD8^(+)TILs and higher CD57^(+)CD8^(+)TALs,assessed prior to treatment,correlated with poor prognosis and suppressive TIME in advanced HGSOC.
