Human Tissue Acquisition and Processing Core

人体组织采集和处理核心（科学核心B）协调采集 从中获得肥大细胞、嗜碱性粒细胞和B细胞的人组织。组织来源的肥大细胞 以及来自外周血的嗜碱性粒细胞和B细胞，当它们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。他们没有染色体畸变， 基因表达异常的白血病细胞，也表现出关键的差异，从那些细胞来源 在体外从祖细胞。例如，脐带血来源的肥大细胞主要表达FcvRIIb， 从皮肤和肺分散的那些仅表达Fc γ Rlla，一种活化受体。 新鲜手术肺和皮肤将用作成熟人肥大细胞的来源，外周血用于 嗜碱性粒细胞和B细胞。MCT细胞是肺中的主要肥大细胞类型，MCTc细胞在皮肤中。的 国家疾病研究交流中心（http：//www.ndriresource.org/），合作人体组织 网络（http：//www-chtn.ims.nci.nih.gov/）和VCD病理科将提供皮肤和肺; 和弗吉尼亚血液服务（http：//www.vablood.org/）（血沉棕黄层细胞）和个人捐献者将 提供外周血来源。将从肺和皮肤中分散肥大细胞，用抗Kit纯化 mAb，根据需要用抗CD 88 Ab分离成MCT和MCTc细胞，置于培养物中并分配 适当的项目。来自血沉棕黄层或外周血的嗜碱性粒细胞和B细胞将通过 密度依赖性沉降和负选择，分布。项目1需要组织来源的 肥大细胞和外周血嗜碱性粒细胞以研究FceRI和/或Fc γ RIIa介导的脱敏，和 促进与Spiegel博士（P4）就SphK和CerK的参与以及与Ryan博士的合作 (P3)林恩参与脱敏治疗的证据项目2利用外周血或血沉棕黄层的B细胞 将CD 23依赖性调节IgE产生的动物模型扩展到人类， 体外新的CD 23调节剂对IgE产生的影响。项目4利用组织来源的 研究鞘氨醇和神经酰胺激酶的新型抑制剂对介质 这些细胞被激活后的生产，并促进与施瓦茨博士的合作， Kepley关于这些酶对人类肥大细胞功能的作用。核心B也将管理奥德赛 红外成像系统，这将主要用于处理和分析蛋白质印迹的 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

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.