CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance...CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.展开更多
目的 检测长链非编码RNA(LncRNA)癌易感性候选基因7(CASC7)、微小RNA-21(miR-21)在支气管哮喘急性发作期患儿外周血中的表达水平并分析其临床意义。方法 选取2020年6月至2021年10月在襄阳市中心医院就诊的支气管哮喘急性发作期患儿115例...目的 检测长链非编码RNA(LncRNA)癌易感性候选基因7(CASC7)、微小RNA-21(miR-21)在支气管哮喘急性发作期患儿外周血中的表达水平并分析其临床意义。方法 选取2020年6月至2021年10月在襄阳市中心医院就诊的支气管哮喘急性发作期患儿115例(急性发作期组)以及支气管哮喘缓解期患儿97例(缓解期组),根据病情程度将急性发作期组患儿分为轻度组45例,中度组39例,重度组31例。另收集同期体检健康儿童100例(健康人对照组)。采用实时荧光定量PCR法(RT-qPCR)检测各组外周血LncRNA CASC7、miR-21的表达水平;检测第1秒用力呼气容积(forced expiratory volume in 1 second, FEV1)、用力肺活量(forced vital capacity, FVC)、最大呼吸峰流速(peak expiratory flow, PEF)等肺功能指标;采用免疫比浊法检测超敏C反应蛋白(hs-CRP)的表达,ELISA法检测TNF-α、IL-6的表达水平;Pearson法分析LncRNA CASC7、miR-21的表达与hs-CRP、TNF-α、IL-6及FEV1、FVC、PEF的相关性,并进一步分析LncRNA CASC7与miR-21的相关性;ROC曲线分析LncRNA CASC7、miR-21对支气管哮喘急性发作期患儿的鉴别诊断价值。结果 RT-qPCR检测结果显示,健康人对照组、急性发作期组、缓解期组患儿外周血LncRNA CASC7的表达水平(1.09±0.18 vs 0.93±0.12 vs 0.65±0.10)依次降低(F=289.447,P<0.05),miR-21的表达水平(1.05±0.14 vs 1.72±0.35 vs 2.64±0.55)依次升高(F=440.557,P<0.05);轻度组、中度组、重度组哮喘患儿外周血LncRNA CASC7的表达水平(0.78±0.09 vs 0.67±0.09 vs 0.44±0.08)依次降低(F=140.168,P<0.05),而miR-21的表达水平(2.34±0.32 vs 2.63±0.53 vs 2.98±0.42)依次升高(F=20.625,P<0.05);Pearson相关性分析显示,支气管哮喘急性发作期患儿外周血LncRNA CASC7与hs-CRP、TNF-α、IL-6的水平呈负相关(r分别为-0.423、-0.375、-0.398,P均<0.05),而与FEV1、FVC、PEF测定值呈正相关(r分别为0.321、0.453、0.441,P均<0.05);miR-21与hs-CRP、TNF-α、IL-6水平呈正相关(r分别�展开更多
基金the National Research Foundation of Korea(NRF-2021M3A9D3026428)the Ministry of Science and ICT of Korea.J.Jeong is a recipient of funding from the Global Ph.D.Fellowship Program(NRF-2019H1A2A1076865)of the National Research Foundation of Korea.
文摘CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.
文摘目的 检测长链非编码RNA(LncRNA)癌易感性候选基因7(CASC7)、微小RNA-21(miR-21)在支气管哮喘急性发作期患儿外周血中的表达水平并分析其临床意义。方法 选取2020年6月至2021年10月在襄阳市中心医院就诊的支气管哮喘急性发作期患儿115例(急性发作期组)以及支气管哮喘缓解期患儿97例(缓解期组),根据病情程度将急性发作期组患儿分为轻度组45例,中度组39例,重度组31例。另收集同期体检健康儿童100例(健康人对照组)。采用实时荧光定量PCR法(RT-qPCR)检测各组外周血LncRNA CASC7、miR-21的表达水平;检测第1秒用力呼气容积(forced expiratory volume in 1 second, FEV1)、用力肺活量(forced vital capacity, FVC)、最大呼吸峰流速(peak expiratory flow, PEF)等肺功能指标;采用免疫比浊法检测超敏C反应蛋白(hs-CRP)的表达,ELISA法检测TNF-α、IL-6的表达水平;Pearson法分析LncRNA CASC7、miR-21的表达与hs-CRP、TNF-α、IL-6及FEV1、FVC、PEF的相关性,并进一步分析LncRNA CASC7与miR-21的相关性;ROC曲线分析LncRNA CASC7、miR-21对支气管哮喘急性发作期患儿的鉴别诊断价值。结果 RT-qPCR检测结果显示,健康人对照组、急性发作期组、缓解期组患儿外周血LncRNA CASC7的表达水平(1.09±0.18 vs 0.93±0.12 vs 0.65±0.10)依次降低(F=289.447,P<0.05),miR-21的表达水平(1.05±0.14 vs 1.72±0.35 vs 2.64±0.55)依次升高(F=440.557,P<0.05);轻度组、中度组、重度组哮喘患儿外周血LncRNA CASC7的表达水平(0.78±0.09 vs 0.67±0.09 vs 0.44±0.08)依次降低(F=140.168,P<0.05),而miR-21的表达水平(2.34±0.32 vs 2.63±0.53 vs 2.98±0.42)依次升高(F=20.625,P<0.05);Pearson相关性分析显示,支气管哮喘急性发作期患儿外周血LncRNA CASC7与hs-CRP、TNF-α、IL-6的水平呈负相关(r分别为-0.423、-0.375、-0.398,P均<0.05),而与FEV1、FVC、PEF测定值呈正相关(r分别为0.321、0.453、0.441,P均<0.05);miR-21与hs-CRP、TNF-α、IL-6水平呈正相关(r分别�
