Human Tissue Acquisition and Processing Core

人体组织采集和处理核心

• 批准号: 7476206
• 负责人: Wei Zhao
• 金额: $ 21.81万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7476206
• 关键词: Animal Model; B-Lymphocytes; Basophils; Blood; Cell Density; Cell physiology; Cells; Chromosome abnormality; Collaborations; Cooperative Human Tissue Network; Disease; Endopeptidases; Environment; Enzymes; Exhibits; Flow Cytometry; Gene Expression; Glycocalyx; Human; IgE; Image; In Vitro; Individual; Label; Leukemic Cell; Lung; Mediating; Mediator of activation protein; Operative Surgical Procedures; Pathology; Peptide Hydrolases; Phosphotransferases; Process; Production; Protein Tyrosine Kinase; Regulation; Research; Research Personnel; Role; Sampling; Schedule; Sedimentation process; Services; Signal Transduction; Skin; Sorting - Cell Movement; Source; Specimen; Sphingosine; Standards of Weights and Measures; System; Techniques; Tissues; Umbilical Cord Blood; Universities; Virginia; Western Blotting; cell type; ceramide kinase; density; desensitization; human tissue; in vivo; inhibitor/antagonist; magnetic beads; mast cell; novel; peripheral blood; progenitor; receptor

The Human Tissue Acquisition and Processing Core (Scientific Core B) coordinates the acquisition of human tissues from which mast cells, basophils and B cells are obtained. Mast cells derived from tissues and basophils and B cells from peripheral blood, after they have matured in their natural environments, best reflect the functional attributes of these cells in vivo. They lack the chromosomal aberrations and gene expression abnormalities of leukemic cells, and also exhibit key differences from those cells derived in vitro from progenitors. For example, cord blood-derived mast cells express primarily FcvRllb, an inhibitory receptor, while those dispersed from skin and lung express only FcyRlla, an activating receptor. Fresh surgical lung and skin will be used as sources of mature human mast cells, peripheral blood for basophils and B cells. The MCT cell is the predominant mast cell type in lung, the MCTc cell in skin. The National Diseases Research Interchange (http://www.ndriresource.org/), Cooperative Human Tissue Network (http://www-chtn.ims.nci.nih.gov/) and Pathology Department at VCD will provide skin and lung; and the Virginia Blood Services (http://www.vablood.org/) (buffy coat cells) and individual donors will provide a source of peripheral blood. Mast cells will be dispersed from lung and skin, purified with anti-Kit mAb, separated into MCT and MCTc cells with anti-CD88 Ab as needed, placed into culture and distributed to the appropriate Projects. Basophils and B cells from buffy coats or peripheral blood will be purified by density-dependent sedimentation and negative selection, and distributed. Project 1 requires tissue-derived mast cells and peripheral blood basophils to study FceRI and/or FcyRlla mediated desensitization, and facilitates collaborations with Dr. Spiegel (P4) on the involvement of SphK and CerK and with Dr. Ryan (P3) on the involvement of Lyn in desensitization. Project 2 utilizes B cells from peripheral blood or buffy coats to extend the animal model of CD23-dependent regulation of IgE production to humans by assessing in vitro the effect of novel CD23 modulating agents on IgE production. Project 4 utilizes tissue-derived mast cells to examine the effects of novel inhibitors of sphingosine and ceramide kinases on mediator production by these cells after they are activated, and facilitates collaborations with Drs. Schwartz and Kepley on the roles of these enzymes on human mast cell functions. Core B also will manage the Odyssey Infrared Imaging System, which will be used primarily for processing and analyzing Western blots of the signal transduction molecules under study in Projects 1-4. Thus, Core B relates to each of the proposed Projects.

人体组织获取和处理核心(科学核心B)协调获取 获得肥大细胞、嗜碱性粒细胞和B细胞的人体组织。来源于组织的肥大细胞 外周血液中的嗜碱性粒细胞和B细胞，在自然环境中成熟后， 最能反映这些细胞在体内的功能属性。它们缺乏染色体的畸变率 白血病细胞的基因表达异常，也显示出与那些来源的细胞的关键差异 来自祖细胞的体外培养。例如，脐带血来源的肥大细胞主要表达FcvRllb，一个 而从皮肤和肺散布的受体只表达激活受体FcyR11a。 新鲜的外科手术肺和皮肤将被用作成熟的人类肥大细胞的来源，外周血用于 嗜碱性粒细胞和B细胞。MCT细胞是肺内最主要的肥大细胞类型，皮肤中以MCTC细胞为主。这个 国家疾病研究交流中心(http://www.ndriresource.org/)，合作人体组织 网络(VCD的http://www-chtn.ims.nci.nih.gov/)和病理科将提供皮肤和肺； 和弗吉尼亚州血液服务局(http://www.vablood.org/)(巴菲外套细胞)和个人献血者将 提供外周血源。肥大细胞将从肺和皮肤中分散，用抗试剂盒纯化 根据需要用抗CD88单抗将单抗分离成MCT和MCTC细胞，放入培养中并分布 到适当的项目。从棕黄色大衣或外周血液中提取的嗜碱性粒细胞和B细胞将通过 依赖密度的沉积和负选择，并呈分布。项目%1需要组织衍生 肥大细胞和外周血嗜碱性粒细胞研究FceRI和/或FcyR11a介导的脱敏，以及 促进与Spiegel博士(P4)就SphK和Cerk的参与以及与Ryan博士的合作 (P3)LYN参与脱敏。项目2利用外周血液或缓冲液中的B细胞 COATS通过评估将CD23依赖的IgE产生调节的动物模型扩展到人类 新型CD23调节剂在体外对IgE产生的影响项目4利用从组织中提取的 肥大细胞检测新型鞘氨醇和神经酰胺酶抑制剂对介质的影响 在激活后由这些细胞产生，并促进与Schwartz博士和 Kepley关于这些酶在人类肥大细胞功能中的作用。核心B也将管理奥德赛 红外成像系统，主要用于处理和分析 项目1-4中正在研究的信号转导分子。因此，核心B涉及每个拟议的 项目。