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鸡FOXP3结构预测及组织表达谱分析

生物工程学报 2022年第8期38卷 2928-2938页

作者：常贺高辉王永强高丽曹红李晓齐郑世军中国农业大学动物医学院农业动物流行病学重点实验室北京100193

本研究旨在克隆鸡FOXP3(chFOXP3)基因全长编码区序列(coding sequence,CDS),并对其进行生物信息学及组织表达谱分析。以50日龄无特定病原鸡为样本,从脾脏组织中克隆chFOXP3基因全长CDS序列,利用在线工具和软件对chFOXP3进行生物信息学分... 详细信息

本研究旨在克隆鸡FOXP3(chFOXP3)基因全长编码区序列(coding sequence,CDS),并对其进行生物信息学及组织表达谱分析。以50日龄无特定病原鸡为样本,从脾脏组织中克隆chFOXP3基因全长CDS序列,利用在线工具和软件对chFOXP3进行生物信息学分析,以实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术检测chFOXP3基因在鸡各组织中的表达分布情况。研究结果表明,chFOXP3基因包含882 bp的开放阅读框,编码293个氨基酸,chFOXP3分子质量为33.44 kDa,属于亲水蛋白;chFOXP3具有Fox转录因子家族典型的forkhead结构域,含有核定位信号;其二级结构由α-螺旋(29.35%)、延伸链(10.92%)、β-转角(5.12%)和无规则卷曲(54.61%)组成;chFOXP3在不同组织中表达有差异,在鸡心脏及胰腺中表达水平较高,显著高于脾脏、法氏囊以及胸腺等免疫器官(PFOXP3与其他野禽属于相同分支,但与哺乳动物相差较远。这些研究结果为进一步深入研究chFOXP3的免疫调节功能奠定了基础。

关键词：鸡 FOXP3 克隆表达分析

DNA bridging of FOXP3 ladder-like multimer: Unveiling a novel transcriptional regulation paradigm

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The Innovation 2024年第3期5卷 15-16页

作者： Ying Lu Lanfang Li Hanting Yang Bin Li Zuoyun Wang Key Laboratory of Metabolism and Molecular Medicine of the Ministry of Education Shanghai Xuhui Central Hospital and School of Basic Medical SciencesFudan UniversityShanghai 200032China Institute for Translational Brain Research State Key Laboratory of Medical NeurobiologyMOE Frontiers Center for Brain ScienceDepartment of NeurologyZhongshan HospitalFudan UniversityShanghai 200032China Center for Immune-Related Diseases at Shanghai Institute of Immunology Department of Immunology and MicrobiologyShanghai Jiao Tong University School of MedicineShanghai 200025China

In the latest issue of Nature,Zhang et al.characterized a novel ladder-like structure of FOXP3-DNA interaction involving FOXP3 multimerization and remote DNA bridging through a combination of biochemistry,structural biology,cell biology,and bioinformatics analyses.1 In this commentary,we highlight their key findings and provide our insights into the research paradigm for further exploration of a novel transcriptional regulation mode as well as a therapeutic strategy from the structural aspects of the FOXP3 complex(Figure 1).

关键词： FOXP3 bridging

Bel 10 is required for the development and suppressive function of FOXP3^(+)regulatory T cells

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Cellular & Molecular Immunology 2021年第1期18卷 206-218页

作者： Dandan Yang Xueqiang Zhao Xin Lin Department of Basic Medical Sciences and Institute for Immunology Tsinghua University School of MedicineBeijing 100084China Tsinghua University-Peking University Jointed Center for Life Sciences Beijing 100084China

FOXP3^(+)regulatory T(Treg)cells play a critical role in peripheral tolerance.Bcl1O,acting as a scaffolding protein in the Carma1-Bcl10-Malt1(CBM)complex,has a critical role in TCR-induced signaling,leading to NF-kB activation and is required for T-cell activation.The role of Bcl1O in conventional T(Tconv)cells has been well characterized;however,the role of Bcl1 in the development of Treg cells and the maintenance of the suppressive function and identity of these cells has not been well characterized.In this study,we found that Bcl10 was required for not only the development but also the function of Treg cells.After deleting Bcl1O in T cells,we found that the development of Treg cells was significantly impaired.When Bcl1O was specifically deleted in mature Treg cells,the suppressive function of the Treg cells was impaired,leading to lethal autoimmunity in Bcl10^(fl/fl)FOXP3^(cre) mice.Consistently,in contrast to WT Treg cells,Bcl10-deficient Treg cells could not protect Rag1-deficient mice from T-cell transfer-induced colitis.Furthermore,Bcl1O-deficient Treg cells downregulated the expression of a series of Treg-cell effector and suppressive genes and decreased effector Treg-cell populations.Moreover,Bcl1O-deficient Treg cells were converted into IFNγ-producing proinflammatory cells with increased expression of the transcription factors T-bet and HIF-1α.Together,our study results provide genetic evidence,indicating that Bcl1O is required for the development and function of Treg cells.

关键词： Bcl1O Treg cells FOXP3 autoimmune inflammation suppressive function

姜黄素调控转录因子FOXP3影响HIV-1感染辅助受体CCR5的作用机制研究

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中国免疫学杂志 2024年第4期40卷 772-779页

作者：冯龙李青雅李寒冰王白燕曹珊郑文锦耿宇轩李青河南中医药大学医学院郑州450046

目的:探讨姜黄素通过调控转录因子FOXP3影响HIV-1感染辅助受体CCR5的作用机制。方法:利用生物信息学方法预测并分析转录因子FOXP3与CCR5启动子的结合位点;采用AutoDock 4.2软件对姜黄素与FOXP3进行柔性对接;MTT法检测姜黄素对Jurkat细... 详细信息

目的:探讨姜黄素通过调控转录因子FOXP3影响HIV-1感染辅助受体CCR5的作用机制。方法:利用生物信息学方法预测并分析转录因子FOXP3与CCR5启动子的结合位点;采用AutoDock 4.2软件对姜黄素与FOXP3进行柔性对接;MTT法检测姜黄素对Jurkat细胞活性的影响;qRT-PCR和Western blot检测不同浓度姜黄素作用于Jurkat细胞后CCR5和FOXP3 mRNA和蛋白表达水平;构建pcDNA3.1-FOXP3真核表达载体;结合转录因子预测结果,运用Overlap PCR法扩增突变型CCR5基因片段,构建突变型CCR5启动子报告载体pFireRluc-Mt-CCR5;利用双荧光素酶报告基因技术验证转录因子FOXP3与CCR5的启动子结合位点。结果:JASPAR转录因子预测结果显示,CCR5启动子区与转录因子FOXP3存在结合位点;分子对接结果显示,姜黄素能够与FOXP3的酶活区域结合;MTT结果显示,姜黄素作用24 h后对Jurkat细胞活性产生抑制作用,IC50为34.48μmol/L;qRT-PCR和Western bot结果显示,不同浓度姜黄素作用于Jurkat细胞后,CCR5和FOXP3 mRNA和蛋白表达水平均降低,且存在剂量依赖性;双荧光素酶报告基因技术证实FOXP3能够与CCR5启动子结合,且转录因子FOXP3可调控CCR5启动子活性;过表达FOXP3后,姜黄素对CCR5的作用结果显示:当姜黄素浓度为60μmol/L时,作用于共转染pcDNA3.1-FOXP3和pFireRluc-Wt-CCR5的HEK293T细胞CCR5启动子荧光素酶活性相对值明显高于pFireRluc-Wt-CCR5+curcumin-60组(PFOXP3能够调控CCR5启动子活性,其作用机制可能是姜黄素通过作用于FOXP3与CCR5启动子结合位点影响CCR5启动子活性。

关键词：姜黄素 FOXP3 CCR5 HIV-1 调控

FOXP3 protects conventional human T cells from premature restimulation-induced cell death

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Cellular & Molecular Immunology 2021年第1期18卷 194-205页

作者： Kelsey Voss Camille Lake Christopher R.Luthers Nathaniel M.Lott Batsukh Dorjbal Swadhinya Arjunaraja Bradly M.Bauman Anthony R.Soltis Gauthaman Sukumar Clifton LDalgard Andrew L.Snow Department of Pharmacology&Molecular Therapeutics Uniformed Services University of the Health SciencesBethesdaMD 20817USA The American Genome Center(TAGC) Collaborative Health Initiative Research ProgramUniformed Services University of the Health SciencesBethesdaMDUSA Department of Anatomy Physiology&GeneticsUniformed Services University of the Health SciencesBethesdaMDUSA

The adaptive immune response relies on specific apoptotic programs to maintain homeostasis.Conventional effector T cell(Tcon)expansion is constrained by both forkhead box P3(FOXP3)^(+)-regulatory T cells(Tregs)and restimulation-induced cell death(RICD),a proprlocidal apoptosis pathway triggered by repeated stimulation through the T-cell receptor(TCR).Constitutive FOXP3 expression protects Tregs from RICD by suppressing SLAM-associated protein(SAP),a key adaptor protein that amplifies TCR signaling strength.The role of transient FOXP3 induction in activated human CD4 and CD8 Tcons remains unresolved,but its expression is inversely correlated with acquired RICD sensitivity.Here,we describe a novel role for FOXP3 In protecting human Tcons from premature RICD during expansion.Unlike FOXP3-mediated protection from RICD in Tregs,FOXP3 protects Tcons through a distinct mechanism requiring de novo transcription that does not require SAP suppression.Transcriptome profiling and functional analyses of expanding Tcons revealed that FOXP3 enhances expression of the SLAM family receptor CD48,which in turn sustains basal autophagy and suppresses pro-apoptotic p53 signaling.Both CD48 and FOXP3 expression reduced p53 accumulation upon TCR restimulation.Furthermore,silencing FOXP3 expression or blocking CD48 decreased the mitochondrial membrane potential in expanding Tcons with a concomitant reduction in basal autophagy.Our findings suggest that FOXP3 governs a distinct transcriptional program in early-stage effector Tcons that maintains RICD resistance via CD48-dependent protective autophagy and p53 suppression.

关键词： conventional T cells RICD FOXP3 autophagy CD48

FOXP3在激素受体阳性乳腺癌中的表达及其对MCF-7细胞侵袭和迁移的影响

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解放军医学杂志 2022年第3期47卷 269-276页

作者：李静平任晓菲刘北辰曹淼任曙光赵小涵刘运江张香梅河北医科大学第四医院乳腺中心石家庄050011 河北省肿瘤微环境与耐药重点实验室石家庄050011 河北医科大学第四医院血液科石家庄050011 河北医科大学第四医院动物中心石家庄050011 河北医科大学第四医院放疗科石家庄050011 河北医科大学第四医院科研中心石家庄050011

目的探讨转录因子FOXP3在激素受体阳性乳腺癌组织中的表达及其对MCF-7细胞侵袭和迁移的影响。方法收集2010年9月-2011年10月河北医科大学第四医院乳腺中心收治的66例激素受体阳性乳腺癌患者的石蜡标本,采用免疫组化检测组织中FOXP3的表... 详细信息

目的探讨转录因子FOXP3在激素受体阳性乳腺癌组织中的表达及其对MCF-7细胞侵袭和迁移的影响。方法收集2010年9月-2011年10月河北医科大学第四医院乳腺中心收治的66例激素受体阳性乳腺癌患者的石蜡标本,采用免疫组化检测组织中FOXP3的表达情况,分析FOXP3表达与临床病理特征的关系;采用Kaplan-Meier法分析FOXP3表达与激素受体阳性乳腺癌预后的关系。采用qRT-PCR检测5种乳腺癌细胞系(MDA-MB-231、MDA-MB-453、MDA-MB-474、BR-3和MCF-7)中FOXP3mRNA的相对表达量,选取FOXP3表达量最高的MCF-7细胞系进行基因沉默。设置si-FOXP3转染组(转染si-FOXP3)、si-NC转染组(转染si-NC)与空白对照组(不做处理),采用Western blotting检测FOXP3蛋白的表达,划痕实验和Transwell实验检测细胞迁移和侵袭能力。结果FOXP3蛋白在激素受体阳性乳腺癌细胞质和细胞核中均有表达,细胞核FOXP3阳性表达率为40.9%(27/66),且在无淋巴结转移、Ki-67低表达、Luminal A型及TNM分期Ⅰ期乳腺癌组织中阳性表达率较高(PFOXP3阳性表达率为54.6%(36/66)。Kaplan-Meier生存分析显示,细胞核FOXP3阳性组总生存率明显高于阴性组[96.30%(26/27)vs.82.05%(32/39),P=0.042],而细胞质FOXP3阳性组与阴性组总生存率差异无统计学意义[88.89%(32/36)vs.86.67%(26/30),P=0.715]。qRT-PCR检测结果显示,MCF-7细胞系FOXP3 mRNA相对表达量最高(PFOXP3后,MCF-7细胞的迁移和侵袭能力均较空白对照组明显增强(PFOXP3在激素受体阳性乳腺癌中的预后意义与表达部位相关,细胞核FOXP3的表达可提高总生存率,但细胞质FOXP3的意义尚不明确。FOXP3在乳腺癌MCF-7细胞系中发挥抑制细胞迁移和侵袭的抑癌作用,可作为激素受体阳性乳腺癌的潜在分子标志物。

关键词：乳腺癌 FOXP3 预后迁移侵袭

A wave of FOXP3^(+) regulatory T cell accumulation in the neonatal liver plays unique roles in maintaining self-tolerance

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Cellular & Molecular Immunology 2020年第5期17卷 507-518页

作者： Mingyang Li Weijia Zhao Yifan Wang Lixue Jin Gaowen Jin Xiuyuan Sun Wei Wang Ke Wang Xi Xu Jie Hao Rong Jin Wenxian Fu Ying Sun Yingjun Chang Xiaojun Huang Xuyu Zhou Hounan Wu Kunshan Zhang Qing Ge Department of Immunology School of Basic Medical SciencesNHC Key Laboratory of Medical ImmunologyPeking UniversityBeijingChina Center for Molecular Metabolism Nanjing University of Science&TechnologyNo.364 Building200 Xiaolinwei StreetNanjing 210094China Pediatric Diabetes Research Center Department of PediatricsUCSDLa JollaCAUSA Department of Immunology School of Basic Medical SciencesCapital Medical UniversityBeijingChina Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation Peking University People’s Hospital&Institute of HematologyBeijing 100044China Institute of Microbiology Chinese Academy of SciencesBeijing 100101China Peking University Medical and Health Analytical Center Peking UniversityBeijing 100191China Translational Stem Cell Research Center Tongji HospitalTongji University School of MedicineShanghai 200065China Department of Integration of Chinese and Western Medicine School of Basic Medical SciencesPeking UniversityBeijing 100191China

Newborn animals require tightly regulated local and systemic immune environments to govern the development and maturation of multiple organs/tissues even though the immune system itself is far from mature during the neonatal period.Regulatory T cells(Tregs)are essential for maintaining immune tolerance/homeostasis and modulating inflammatory responses.The features of Tregs in the neonatal liver under steady-state conditions are not well understood.The present study aimed to investigate the phenotype,functions,and significance of neonatal Tregs in the liver.We found a wave of thymus-derived Treg influx into the liver during 1–2 weeks of age.Depletion of these Tregs between days 7 and 11 after birth rapidly resulted in Th1-type liver inflammation and metabolic disorder.More Tregs in the neonatal liver than in the spleen underwent MHC II-dependent activation and proliferation,and the liver Tregs acquired stronger suppressive functions.The transcriptomic profile of these neonatal liver Tregs showed elevated expression of PPARγand T-bet and features of Tregs that utilize lipid metabolic machinery and are capable of regulating Th1 responses.The accumulation of Tregs with unique features in the neonatal liver is critical to ensure self-tolerance and liver maturation.

关键词： Neonatal period liver FOXP3 Treg cells Th1-type inflammation

Treg、FOXP3和TGF-β在慢性髓系白血病中的表达及意义

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中国实验血液学杂志 2023年第3期31卷 666-670页

作者：王舒莉董巧凤李芳王静桑玉旗张琳山东省菏泽市立医院血液内科山东菏泽274031 山东省菏泽市立医院检验科山东菏泽274031

目的:分析调节性T细胞(Treg)、叉头状转录因子P3(FOXP3)和转化生长因子-β(TGF-β)在慢性髓系白血病(CML)不同时期中的表达及意义。方法:收集2018年3月-2021年3月在菏泽市立医院血液内科就诊的73例CML患者的外周血,根据患者所处的CML不... 详细信息

目的:分析调节性T细胞(Treg)、叉头状转录因子P3(FOXP3)和转化生长因子-β(TGF-β)在慢性髓系白血病(CML)不同时期中的表达及意义。方法:收集2018年3月-2021年3月在菏泽市立医院血液内科就诊的73例CML患者的外周血,根据患者所处的CML不同时期,将患者分为初诊组、慢性期组和急变期组,应用流式细胞术检测患者Treg水平,RT-q PCR检测FOXP3 mRNA表达水平,酶联免疫吸附试验检测血浆中TGF-β的表达水平,分析上述指标在患者临床发病过程中的作用。结果:初诊组患者Treg比例较慢性期组升高,但差异无显著统计学意义(P=0.695),而急变期组较初诊组和慢性期组均显著升高(P=0.008,PFOXP3mRNA表达水平分别为11.61±2.21、6.46±1.35和8.54±2.13,FOXP3 mRNA表达水平在CML不同时期的差异具有统计学意义(F=55.199,PFOXP3 mRNA表达水平均高于慢性期组(P34,P=0.728)。结论:Treg细胞亚群在CML不同时期中的异常分布和FOXP3 mRNA在CML初诊时和急变期的表达水平显著升高,表明Treg亚群可能通过免疫调节促进CML的发生和进展。

关键词： FOXP3 调节性T细胞慢性髓系白血病

非小细胞肺癌血浆天然CD25抗体及FOXP3抗体的表达变化及其作为诊断标志物的探讨

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中国免疫学杂志 2022年第1期38卷 80-85页

作者：赵欢张萱李庆海(指导) 青岛大学附属青岛市市立医院内分泌科青岛266071 吉林大学第二医院吉林省分子与遗传学重点实验室长春130031 青岛大学附属青岛市市立医院呼吸科青岛266071

目的:探讨天然CD25和FOXP3抗体在非小细胞肺癌(NSCLC)患者血浆表达变化的临床意义及其作为诊断标志物的潜能。方法:选取青岛大学附属青岛市立医院胸外科首次确诊且未经任何治疗的NSCLC患者200例及同期性别、年龄、吸烟史相匹配的健康志... 详细信息

目的:探讨天然CD25和FOXP3抗体在非小细胞肺癌(NSCLC)患者血浆表达变化的临床意义及其作为诊断标志物的潜能。方法:选取青岛大学附属青岛市立医院胸外科首次确诊且未经任何治疗的NSCLC患者200例及同期性别、年龄、吸烟史相匹配的健康志愿者190例,收集两组血液样本。基于CD25及FOXP3分子结构,利用表位预测软件(http://***)设计并合成3条线性抗原肽(即CD25a、CD25b、FOXP3)并外送化工公司合成,通过自行建立的优化ELISA检测方法,检测NSCLC患者及健康对照组血浆CD25及FOXP3抗体的表达水平,并分析CD25及FOXP3抗体的表达与临床病理特征之间的关系。同时绘制ROC曲线,评估CD25及FOXP3抗体对不同TNM分期NSCLC的诊断价值。结果:NSCLCⅡ~Ⅳ期患者血浆CD25a、FOXP3抗体的表达较健康对照组显著增加(0.66±0.18 vs 0.52±0.17,P3 vs 0.53±0.25,P37±0.21 vs 0.43±0.22,P0.05)。CD25b抗体的表达与性别、年龄、肿瘤分化程度有关(P均0.05),而FOXP3抗体的表达仅与肿瘤分化程度有关(PFOXP3抗体诊断NSCLC的效能评估,CD25a曲线下面积为0.727(95%CI:0.677~0.778),最佳截断值为0.55,敏感度为77%,特异度为64%;CD25b曲线下面积为0.601(95%CI:0.545~0.657),最佳截断值为0.35,敏感度为62%,特异度为57%;FOXP3曲线下面积为0.613(95%CI:0.557~0.669),最佳截断值为0.54,敏感度为58%,特异度为65%。结论:检测CD25a天然抗体可辅助NSCLC的早期诊断;血浆CD25b天然抗体在早期NSCLC(Ⅰ+Ⅱ期)表达降低,对预测早期NSCLC有一定价值。

关键词：天然抗体 CD25 FOXP3 非小细胞肺癌