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Applications of cytof in Brain Immune Component Studies

Engineering 2022年第9期16卷 187-197页

作者： Yan Wang Baohui Xu Lixiang Xue Department of Neurosurgery Peking University Third HospitalBeijing 100191China Medical Research Center Peking University Third HospitalBeijing 100191China Center of Basic Medical Research Institute of Medical Innovation and ResearchPeking University Third HospitalBeijing 100191China Department of Vascular Surgery Stanford University School of MedicineStanfordCA 94305USA Biobank Peking University Third HospitalBeijing 100191China

The brain is the most heterogeneous and complex tissue in the body.Previous studies have shown that immune cells are essential functional components in both healthy and pathological brains.Cytometry by the time of flight(cytof)is a high-dimensional single-cell detection technology that allows measurements of up to 100 cell markers with a small number of samples.This technique enables the identification and characterization of various cell types at the single-cell level under steady-state and diseased brain conditions.This review outlines three major advantages of the cytof technique compared with the traditional flow cytometry approach.We also discuss cytof applications in brain immune cell component research in both healthy and pathological brains.

关键词： cytof Brain Immune components Neuroinflammation

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Advanced technologies for single-cell in situ protein profiling

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Science China Chemistry 2022年第1期65卷 48-67页

作者： Wen Zhou Xiaoyue Ni Chen Xie Quli Fan Dingbin Liu State Key Laboratory of Organic Electronics and Information Displays and Jiangsu National Synergetic Innovation Center for Advanced Materials(SICAM)Institute of Advanced Materials(IAM)Nanjing University of Posts&TelecommunicationsNanjing 210023China State Key Laboratory of Medicinal Chemical Biology Research Center for Analytical SciencesTianjin Key Laboratory of Biosensing and Molecular RecognitionCollege of ChemistryNankai UniversityTianjin 300071China

Cells are inherently heterogeneous to achieve a diverse spectrum of biological functions.To understand the underlying protein machinery that achieves these fascinating functions,it is important to develop advanced analytical methods that can profile proteins in their native environment,as protein expression,aggregation,degradation,and regulation define both normal physiology as well as pathogenicity.Genome and transcriptome sequencing have seen major advances at the single-cell levels,but comprehensive proteomic profiling is still challenging.The conventional proteomic methods,such as enzyme-linked immunosorbent assay(ELISA),western blot,and protein chips,can characterize biomarkers of interest.Still,these ensemble techniques are unsuitable for single-cell studies.Increasing evidence has shown the significance of in situ,sensitive,quantitative,and multiplexed profiling of biomarkers in single cells for diagnosis and treatment guidance.Here,we review the recent development of advanced imaging and spectroscopy techniques,including mass cytometry,immunofluorescence,and surfaceenhanced Raman spectrometry(SERS)for single-cell proteomic imaging.We also provide our view on the challenges and the outlook.

关键词： proteomics single-cell cytof immunofluorescence SERS

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T淋巴细胞相关亚群变化在急性胰腺炎疾病发展中的意义

T淋巴细胞相关亚群变化在急性胰腺炎疾病发展中的意义

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作者：曾华火福建医科大学

学位级别：硕士

目的:研究T淋巴细胞相关亚群在急性胰腺炎发病过程中的免疫表征变化并探讨其意义。方法:设置急性胰腺炎(AP)组及健康对照组。AP组收录5例符合急性胰腺炎诊断标准的胰腺炎患者,收集患者信息,每名患者分别采集发病后第一天、第三天、第七... 详细信息

目的:研究T淋巴细胞相关亚群在急性胰腺炎发病过程中的免疫表征变化并探讨其意义。方法:设置急性胰腺炎(AP)组及健康对照组。AP组收录5例符合急性胰腺炎诊断标准的胰腺炎患者,收集患者信息,每名患者分别采集发病后第一天、第三天、第七天这三个时间点的外周血样本10ml,进行离心后去上清收集全血细胞。健康对照组收录同时期5例健康体检人员,抽取健康组5例人员外周血液样本10ml进行离心后去上清收集全血细胞。共得到血液样本共20管,血液样本通过Ficoll密度梯度离心法分离出PBMC(外周血单个核细胞)后继续制成单细胞悬液后,选择42个免疫相关点位或者受体,进行细胞内外相关marker的染色后,于Cy TOF(质谱细胞流式技术)仪器上收集相关数据,用Xshift聚类分析出相关联的亚群类别,MST可视化及vi SNE散点图进行分析各类亚群、各maker表达情况,深度分析胰腺炎患者不同时间点的、以T淋巴细胞为主的免疫表达差异,通过t检验对所有亚群变化进行统计分析,找出变化有意义的亚群,分析有变化意义的亚群细胞的比例变化、代谢特征、效应因子及共抑制分子的表达情况。结果:1.急性胰腺炎组病人与健康对照组病人相比,血清AMS、CRP、中性粒细胞表达均升高,具有显著差异。2.T细胞相关亚群表达情况:急性胰腺炎组病人与健康对照组病人,其外周血中有10组淋巴细胞亚群的表达具有显著变化:(1)亚群C22、C17(中性粒细胞亚群)、C26(Fox P3+,CD152-CTLA-4+RORr+,CD154-CD40L+的嗜酸性粒细胞亚群),在急性胰腺炎的疾病发展中较健康对照组表达明显上升。(2)C13(单核巨噬细胞亚群)、C14(CD38+,CD39+的常规树突状细胞亚群)、C27(嗜酸性粒细胞亚群)、C28(CD8a+初始T细胞)、C31(CD8+效应记忆T细胞)、C34(CD8+,h-m-GranzymeB+效应记忆T细胞)、C38(CD4+效应记忆T细胞)在急性胰腺炎的疾病发展中较健康对照组表达明显下降。差异都具有统计学意义(P<0.05)。3、健康对照组和急性胰腺炎组中差异表达的16种marker:CD27,CD14,Lag-3,CD39,PD-L1,CD59,Gata3-h/m,CD127,CTLA-4,FoxP3,CD137,RORy,CD154,h/m-Granzyme B,HLA-DR,CD8a的免疫特征(p<0.05,符合同一分布的概率,普遍意义上认为在一次实验中,这样的样本组是真的有差异)。其中CD27,CD39,HLA-DR,CD8a,CD127较健康对照组差异性下降,CD14,Lag-3,CD39,PD-L1,Gata3-h/m,h/m-Granzyme B,CD154,RORy,CD137,FoxP3,CTLA-4较健康组差异性上升。共抑制因子PD1、CD28、Tim-3在急性胰腺炎疾病发展中无明显表达。结论:1、急性胰腺炎的免疫抑制可能与CD38+,CD39+的常规树突状细胞亚群、CD8a+初始T细胞亚群、CD8+效应记忆T细胞、CD8+,h-m-Granzyme B+效应记忆T细胞亚群、CD4+效应记忆T细胞亚群的细胞占比下降有关。2、CD14,Lag-3,CD39,PD-L1,Gata3-h/m,h/m-Granzyme B,CD154,RORy,CD137,Fox P3,CTLA-4在急性胰腺炎发展中表达量呈现显著性上升,预示其可能被用作急性胰腺炎的潜在生物标志物。CD27,CD39,HLA-DR,CD8a,CD127在急性胰腺炎发展中显著性下降,预示其可能是急性胰腺炎潜在的提示预后指标或者治疗靶点。3、共抑制因子PD1、CD28、Tim-3在急性胰腺炎疾病发展中无明显表达。

关键词：急性胰腺炎 T细胞亚群 cytof

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Deep Profiling of B-cell acute lymphoblastic leukemia with 36-Parameter Mass Cytometry

Deep Profiling of B-cell acute lymphoblastic leukemia with 3...

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第十二届全国免疫学学术大会

作者： Jia Xian Li Jian Huang Jinqi Chen Xian Chen Xiaolei Xiao Changchun Fu Guo Xiamen University

Heterogeneity has long been recognized as a key feature of acute lymphocytic leukemia(ALL)and may lead to the differential responses of ALL patients to therapy.A comprehensive understanding of the immunophenotypes of ... 详细信息

关键词： B-cell acute lymphoblastic leukemia Mass Cytometry cytof immunophenotype T cells

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Estrogen inhibits the growth of colon cancer in mice through reversing extracellular vesicles-mediated immunosuppressive tumor microenvironment

Estrogen inhibits the growth of colon cancer in mice through...

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第十四届全国免疫学学术大会

作者： Lingling Jiang Haiyi Fei Songying Zhang Sir Run Run Shaw Hospital School of MedicineZhejiang University

Postmenopausal women taking estrogen supplements have a lower risk of advanced colorectal cancer,but the exact mechanism involved remains unclear.Thus,this study examined the role of estrogen in colorectal cancer.The estrogen receptor expression in in situ colorectal cancer tissue from female patients increased significantly,indicating their estrogen sensitivity.Compared with the sham-operated group,the growth of MC38 tumors was enhanced in ovariectomized mice,which was canceled in ovariectomized mice with E2 supplementation.The population of PD-L1M2-like macrophages,regulatory T(Treg) cells and myeloid-derived suppressor cells(MDSCs) significantly increased,and the population of cytotoxic CD8T cells declined in MC38 tumors in ovariectomized mice,which were all reversed in ovariectomized mice with E2 supplementation.Extracellular vesicles derived from MC38 cells(MC38-EVs) were involved in the establishment of immunosuppressive tumor microenvironment,but not EVs derived from MC38 cells treated with E2(E2-MC38-EVs),which had reduced level of TGF-β1 and were less capable of inducing Treg in vitro comparing with MC38-EVs.Overall,these results revealed that estrogen restraint MC38 tumor growth via regulating the tumor immune microenvironment,which was mediated by MC38-EVs.This study provides a rationale for further studies on postmenopausal female patients,and suggest that combining estrogen replacement treatment with other anti-tumor therapies may be beneficial in preventing or limiting the outgrowth of colorectal cancer in situ.

关键词： Hormone Gastrointestinal cancer Female Menopause cytof

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Systematical evaluation of SLE patients' immune dysregulation for personalized clinical treatment

Systematical evaluation of SLE patients' immune dysregulatio...

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第十三届全国免疫学学术大会

作者： Lulu zhang Xianting Ding Shanghai Jiao Tong University

Systemic Lupus Erythematosus(SLE) is a highly morbid autoimmune disease characterized by heterogeneous clinical presentation and unpredictable disease activity and the current clinical diagnosis mainly focused on the clinical symptoms rather than related cellular and molecular levels, which limited the development of accurate and personalized treatment for SLE patients.Mass cytometry(cytof) has been widely used for characterization of complex cellular events in terms of its quantify multiple biomarkers in the single-cell suspensions.Here, our study was to evaluate the immune dysregulation of SLE patients systematically with cytof in order to provide some references for clinical application.Herein, we utilized cytof to quantify 27 parameters including phenotypic and functional biomarkers of peripheral blood mononuclear cell(PBMCs) from SLE patients and healthy controls with cytof, respectively.The data were analyzed using the unsupervised hierarchical clustering algorithm ViSNE and SPADE.Our results showed the proportion of CD27 Tc cell of SLE patients had a significant increase compared to the healthy controls, while the na?ve Tc2 cells were variable among all SLE patients.Of note, we can divide the SLE patients into 2 subgroups based on the number of memory B cells, which might provide some information for clinical diagnosis.Altogether, mass cytometry might be a potential tool to promote personalized treatment for SLE patients through analyzing immune system and clinical symptoms.

关键词： SLE cytof Tc2 cells Memory B cells

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