Kang et al.published a research article on the treatment of ischemic stroke using engineered Treg cells(Kang et al.,Prog Biochem Biophys,2025,52(4):946-956.DOI:10.16476/j.pibb.2025.0019).Their study mainly explores th...Kang et al.published a research article on the treatment of ischemic stroke using engineered Treg cells(Kang et al.,Prog Biochem Biophys,2025,52(4):946-956.DOI:10.16476/j.pibb.2025.0019).Their study mainly explores the immunoregulatory role of regulatory T(Treg)cells in ischemic stroke,providing an innovative therapeutic strategy.Neuroinflammation is a major driver of secondary injury after stroke.Existing treatments focus on vascular recanalization while neglecting immune regulation.Their study proposes to modulate neuroinflammation through in vitro-induced Treg cells,offering a novel approach distinct from traditional thrombolysis and endovascular interventions.展开更多
AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiati...AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.展开更多
目的锌指蛋白335(Zfp335)参与调控胸腺T细胞的早期发育和外周T细胞亚群的分化,本研究旨在探讨Zfp335调控调节性T细胞(Treg)在肿瘤免疫中的作用和机制。方法用他莫昔芬在Treg中特异性敲除Zfp335基因[Zfp335^(fl/fl)叉头盒P3(FOXP3)creERT...目的锌指蛋白335(Zfp335)参与调控胸腺T细胞的早期发育和外周T细胞亚群的分化,本研究旨在探讨Zfp335调控调节性T细胞(Treg)在肿瘤免疫中的作用和机制。方法用他莫昔芬在Treg中特异性敲除Zfp335基因[Zfp335^(fl/fl)叉头盒P3(FOXP3)creERT2],并构建MC38移植瘤模型。接种肿瘤后第7天,观察并测量肿瘤的大小,第12天剥离肿瘤组织,用流式细胞术检测野生型(WT)组和Zfp335敲除(Zfp335^(CKO))组小鼠肿瘤浸润淋巴细胞中CD4^(+)T细胞、CD8^(+)T细胞和Treg的比例,以及效应性Treg(eTreg)的线粒体功能。结果自接种肿瘤后第10天开始,Zfp335^(CKO)组肿瘤体积显著小于WT组。Zfp335^(CKO)组CD4^(+)T细胞和CD8^(+)T细胞的肿瘤浸润比例、及其对应的效应细胞比例显著高于WT组。Zfp335^(CKO)组CD4^(+)T细胞和CD8^(+)T细胞分泌细胞因子γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)的比例显著高于WT组,Zfp335^(CKO)组CD8^(+)T细胞分泌颗粒酶B(GzmB)的比例显著高于WT组。Zfp335^(CKO)组Treg、诱导性共刺激分子(ICOS)+Treg比例显著低于WT组。Zfp335^(CKO)组eTreg表达Mitotracker Deep Red的水平显著低于WT组。结论在肿瘤发生过程中,Treg特异性缺失Zfp335导致其活化减弱,与eTreg的线粒体功能降低有关;Zfp335^(CKO)小鼠肿瘤浸润的效应性T细胞增多、分泌杀伤性细胞因子增多,进而抵抗肿瘤发展。展开更多
文摘Kang et al.published a research article on the treatment of ischemic stroke using engineered Treg cells(Kang et al.,Prog Biochem Biophys,2025,52(4):946-956.DOI:10.16476/j.pibb.2025.0019).Their study mainly explores the immunoregulatory role of regulatory T(Treg)cells in ischemic stroke,providing an innovative therapeutic strategy.Neuroinflammation is a major driver of secondary injury after stroke.Existing treatments focus on vascular recanalization while neglecting immune regulation.Their study proposes to modulate neuroinflammation through in vitro-induced Treg cells,offering a novel approach distinct from traditional thrombolysis and endovascular interventions.
基金Supported by the National Natural Science Foundation of China(No.81800423)the Guangdong Medical Science and Technology Research project(No.B2022102)。
文摘AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.
文摘目的锌指蛋白335(Zfp335)参与调控胸腺T细胞的早期发育和外周T细胞亚群的分化,本研究旨在探讨Zfp335调控调节性T细胞(Treg)在肿瘤免疫中的作用和机制。方法用他莫昔芬在Treg中特异性敲除Zfp335基因[Zfp335^(fl/fl)叉头盒P3(FOXP3)creERT2],并构建MC38移植瘤模型。接种肿瘤后第7天,观察并测量肿瘤的大小,第12天剥离肿瘤组织,用流式细胞术检测野生型(WT)组和Zfp335敲除(Zfp335^(CKO))组小鼠肿瘤浸润淋巴细胞中CD4^(+)T细胞、CD8^(+)T细胞和Treg的比例,以及效应性Treg(eTreg)的线粒体功能。结果自接种肿瘤后第10天开始,Zfp335^(CKO)组肿瘤体积显著小于WT组。Zfp335^(CKO)组CD4^(+)T细胞和CD8^(+)T细胞的肿瘤浸润比例、及其对应的效应细胞比例显著高于WT组。Zfp335^(CKO)组CD4^(+)T细胞和CD8^(+)T细胞分泌细胞因子γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)的比例显著高于WT组,Zfp335^(CKO)组CD8^(+)T细胞分泌颗粒酶B(GzmB)的比例显著高于WT组。Zfp335^(CKO)组Treg、诱导性共刺激分子(ICOS)+Treg比例显著低于WT组。Zfp335^(CKO)组eTreg表达Mitotracker Deep Red的水平显著低于WT组。结论在肿瘤发生过程中,Treg特异性缺失Zfp335导致其活化减弱,与eTreg的线粒体功能降低有关;Zfp335^(CKO)小鼠肿瘤浸润的效应性T细胞增多、分泌杀伤性细胞因子增多,进而抵抗肿瘤发展。
文摘目的:分析趋化因子受体8(C⁃C motif chemokine receptor 8,CCR8)在卵巢癌肿瘤浸润性调节性T细胞(regulatory T cell,Treg)中的表达,探讨CCR8对Treg分化的作用。方法:构建C57BL/6小鼠卵巢癌细胞ID8荷瘤模型;流式细胞术检测小鼠肿瘤组织、脾脏和外周血中Treg上CCR8的表达比例,CCR8^(+)Treg上免疫检查点相关蛋白程序性细胞死亡蛋白1(programmed cell death protein 1,PD⁃1)、细胞素性T淋巴细胞抗原4(cytotoxic T⁃lymphocyte antigen 4,CTLA⁃4)、可诱导的T细胞共刺激分子(inducible T cell costimulators,ICOS)、淋巴细胞激活基因3(lymphocyte activation gene 3,LAG⁃3)的表达;流式细胞术检测CCR8变构抑制剂AZ084加入前后对C57BL/6小鼠脾脏中初始CD4^(+)T细胞向Treg分化的影响。结果:卵巢癌荷瘤小鼠肿瘤中Treg上的CCR8表达相比脾脏、外周血的Treg显著增高;相比CCR8^(-)Treg,CCR8^(+)Treg上免疫检查点相关蛋白表达更高;AZ084有效抑制小鼠脾脏中初始CD4^(+)T细胞向Treg的分化。结论:CCR8^(+)Treg在肿瘤浸润性Treg中占主要比例,CCR8作为卵巢癌浸润性Treg的主要标志物,变构CCR8蛋白可以抑制Treg的分化。靶向消除CCR8^(+)Treg可为改善卵巢癌肿瘤微环境的免疫抑制状态提供新思路。
