Leaves and glumes act as lateral organs and have essential effects on photosynthesis and seed morphology,thus affecting yield.However,the molecular mechanisms controlling their polarity development in rice still need ...Leaves and glumes act as lateral organs and have essential effects on photosynthesis and seed morphology,thus affecting yield.However,the molecular mechanisms controlling their polarity development in rice still need further study.Here,we isolated a polarity defect of lateral organs 1(pdl1)mutant in rice,which exhibits twisted/filamentous-shaped leaves and cracked/filamentous-shaped lemmas caused by defects in polarity development.PDL1 encodes a SUPPRESSOR OF GENE SILENCING 3 protein localized in the cytoplasmic granules.PDL1 is expressed in the shoot apical meristem,inflorescence meristem,floral meristem,and lateral organs including leaves and floral organs.PDL1 is involved in the synthesis of tasiR-ARF,which may subsequently modulate the expression of OsARFs.Meanwhile,the expression levels of abaxial miR165/166 and the adaxial identity genes OSHBs were respectively increased and reduced significantly.The results of this study clarify the molecular mechanism by which PDL1-mediated tasiR-ARF synthesis regulates the lateral organ polarity development in rice.展开更多
【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少...【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少PCV2感染造成的经济损失。【方法】利用实验室构建的重组表达宿主菌pET32a-PD1/Rosetta(DE)进行诱导表达、纯化,获得猪可溶性PD-1蛋白(soluble PD-1,sPD-1)的诱导表达条件;制备大量具有活性的猪sPD-1蛋白,体外作用于PCV2感染的猪外周血单个核细胞(peripheral blood mononuclear cell,PBMCs),分别通过CCK-8和流式细胞术检测PBMCs的增殖情况,间接免疫荧光法和RT-qPCR检测猪sPD-1蛋白对PCV2病毒载量的影响,RT-qPCR和ELISA检测猪sPD-1蛋白对免疫相关细胞因子IL-2、IL-12、IL-21、IL-17A和IFN-γ等的转录水平和分泌水平的影响。【结果】通过诱导表达、纯化获得了高纯度、高活性的重组猪sPD-1蛋白。与未使用猪sPD-1蛋白处理的PCV2感染PBMCs组相比,10μg·mL−1猪sPD-1蛋白处理的猪PCV2病毒载量下降至1000 copies·μL−1以下;CCK-8检测结果显示猪sPD-1蛋白处理组细胞增殖指数显著提高(P<0.05);流式细胞术检测发现猪sPD-1蛋白组的平均荧光强度为(68.60±10.14)%,相对于PCV2病毒组(28.70±3.18)%显著增强(P<0.05);反转录荧光定量PCR和ELISA检测,发现猪sPD-1蛋白处理组的细胞因子IL-2、IL-12和IFN-γ的转录水平和分泌水平显著升高(P<0.05)。【结论】猪sPD-1蛋白可以通过阻断PD-1/PD-Ls通路降低PCV2的病毒载量,促进PBMCs的增殖,增强PBMCs细胞因子的转录水平和分泌水平免疫反应。因此,猪sPD-1蛋白在体外可以增强PCV2感染PBMCs的免疫反应,进一步为病毒性疾病的防控提供理论依据。展开更多
目的探讨抗逆转录病毒治疗(antiretroviral treatment,ART)对HIV-1感染者(people living with HIV-1,PLWH)CD3^(+)T细胞PD-1和CD160表达及其活化凋亡水平的影响。方法收集66例慢性HIV-1感染患者,根据是否接受ART治疗,将PLWH分为non-ART...目的探讨抗逆转录病毒治疗(antiretroviral treatment,ART)对HIV-1感染者(people living with HIV-1,PLWH)CD3^(+)T细胞PD-1和CD160表达及其活化凋亡水平的影响。方法收集66例慢性HIV-1感染患者,根据是否接受ART治疗,将PLWH分为non-ART组(39例)和ART组(27例),并设置HIV-1阴性对照组(30例)。采用流式细胞术检测3组外周血CD4计数及CD3^(+)T细胞中PD-1、CD160、CD95、CD38和HLA-DR的表达,并分析3组间不同指标的差异性。结果non-ART组CD3^(+)T细胞百分比和PD-1^(+)T细胞百分比与ART组相比差异无统计学意义(P=0.8141,P>0.9999),而其CD160^(+)CD3^(+)T细胞百分比显著高于ART组(P=0.0162)。共表达分析显示,non-ART组PD-1^(+)CD160^(+)CD3^(+)T细胞百分比显著高于ART组(P=0.0121),而在单阳的CD3^(+)T性细胞(CD160^(+)PD-1^(-)和PD-1^(+)CD160-)细胞群,ART组和non-ART组差异均无统计学意义(P=0.3382,P>0.9999)。在PD-1^(+)CD160^(+)CD3^(+)T细胞CD38方面,non-ART组显著高于ART组和HIV-1阴性对照组(P=0.0005,P=0.0015),HIV-1阴性对照组和ART组差异无统计学意义(P>0.9999)。结论ART对PLWH中PD-1^(+)CD160^(+)CD3^(+)T细胞百分比及其CD38的表达具有调节作用。展开更多
基金supported by the National Natural Science Foundation of China(32470354,31900612,and 31971919)the Natural Science Foundation of Chongqing,China(cstc2020jcyj-jqX0020)+1 种基金the Foundation for Innovative Research Groups of the Natural Science Foundation of Chongqing,China(cstc2021jcyj-cxttX0004)the Chongqing Talent Program Foundation,China(cstc2024ycjhbgzxm0063)。
文摘Leaves and glumes act as lateral organs and have essential effects on photosynthesis and seed morphology,thus affecting yield.However,the molecular mechanisms controlling their polarity development in rice still need further study.Here,we isolated a polarity defect of lateral organs 1(pdl1)mutant in rice,which exhibits twisted/filamentous-shaped leaves and cracked/filamentous-shaped lemmas caused by defects in polarity development.PDL1 encodes a SUPPRESSOR OF GENE SILENCING 3 protein localized in the cytoplasmic granules.PDL1 is expressed in the shoot apical meristem,inflorescence meristem,floral meristem,and lateral organs including leaves and floral organs.PDL1 is involved in the synthesis of tasiR-ARF,which may subsequently modulate the expression of OsARFs.Meanwhile,the expression levels of abaxial miR165/166 and the adaxial identity genes OSHBs were respectively increased and reduced significantly.The results of this study clarify the molecular mechanism by which PDL1-mediated tasiR-ARF synthesis regulates the lateral organ polarity development in rice.
文摘【目的】通过研究猪圆环病毒Ⅱ型(porcine circovirus type Ⅱ,PCV2)感染过程中程序性死亡受体1(programmed cell death protein 1,PD-1)及其配体(programmed cell death 1 ligands,PD-Ls)信号通路作用,寻找调节PCV2免疫抑制新途径,减少PCV2感染造成的经济损失。【方法】利用实验室构建的重组表达宿主菌pET32a-PD1/Rosetta(DE)进行诱导表达、纯化,获得猪可溶性PD-1蛋白(soluble PD-1,sPD-1)的诱导表达条件;制备大量具有活性的猪sPD-1蛋白,体外作用于PCV2感染的猪外周血单个核细胞(peripheral blood mononuclear cell,PBMCs),分别通过CCK-8和流式细胞术检测PBMCs的增殖情况,间接免疫荧光法和RT-qPCR检测猪sPD-1蛋白对PCV2病毒载量的影响,RT-qPCR和ELISA检测猪sPD-1蛋白对免疫相关细胞因子IL-2、IL-12、IL-21、IL-17A和IFN-γ等的转录水平和分泌水平的影响。【结果】通过诱导表达、纯化获得了高纯度、高活性的重组猪sPD-1蛋白。与未使用猪sPD-1蛋白处理的PCV2感染PBMCs组相比,10μg·mL−1猪sPD-1蛋白处理的猪PCV2病毒载量下降至1000 copies·μL−1以下;CCK-8检测结果显示猪sPD-1蛋白处理组细胞增殖指数显著提高(P<0.05);流式细胞术检测发现猪sPD-1蛋白组的平均荧光强度为(68.60±10.14)%,相对于PCV2病毒组(28.70±3.18)%显著增强(P<0.05);反转录荧光定量PCR和ELISA检测,发现猪sPD-1蛋白处理组的细胞因子IL-2、IL-12和IFN-γ的转录水平和分泌水平显著升高(P<0.05)。【结论】猪sPD-1蛋白可以通过阻断PD-1/PD-Ls通路降低PCV2的病毒载量,促进PBMCs的增殖,增强PBMCs细胞因子的转录水平和分泌水平免疫反应。因此,猪sPD-1蛋白在体外可以增强PCV2感染PBMCs的免疫反应,进一步为病毒性疾病的防控提供理论依据。
文摘目的探讨抗逆转录病毒治疗(antiretroviral treatment,ART)对HIV-1感染者(people living with HIV-1,PLWH)CD3^(+)T细胞PD-1和CD160表达及其活化凋亡水平的影响。方法收集66例慢性HIV-1感染患者,根据是否接受ART治疗,将PLWH分为non-ART组(39例)和ART组(27例),并设置HIV-1阴性对照组(30例)。采用流式细胞术检测3组外周血CD4计数及CD3^(+)T细胞中PD-1、CD160、CD95、CD38和HLA-DR的表达,并分析3组间不同指标的差异性。结果non-ART组CD3^(+)T细胞百分比和PD-1^(+)T细胞百分比与ART组相比差异无统计学意义(P=0.8141,P>0.9999),而其CD160^(+)CD3^(+)T细胞百分比显著高于ART组(P=0.0162)。共表达分析显示,non-ART组PD-1^(+)CD160^(+)CD3^(+)T细胞百分比显著高于ART组(P=0.0121),而在单阳的CD3^(+)T性细胞(CD160^(+)PD-1^(-)和PD-1^(+)CD160-)细胞群,ART组和non-ART组差异均无统计学意义(P=0.3382,P>0.9999)。在PD-1^(+)CD160^(+)CD3^(+)T细胞CD38方面,non-ART组显著高于ART组和HIV-1阴性对照组(P=0.0005,P=0.0015),HIV-1阴性对照组和ART组差异无统计学意义(P>0.9999)。结论ART对PLWH中PD-1^(+)CD160^(+)CD3^(+)T细胞百分比及其CD38的表达具有调节作用。
