FLOW CYTOMETRY CORE

核心A包含Becton-Dickinson DiVa细胞分选仪和LSR分析仪。DiVa是一种13参数4向 5-激光细胞分类器。LSR在紫外、可见和红色光谱中具有激发线。核心领导 最近（2005年1月1日）获得了一个SIG，用于建造一个6激光器、20探测器的LSR-2， 程序.对于项目1（Sykes），核心A将用于评估所需的9色流式细胞术研究 GVH反应性和非反应性T细胞亚群上的激活和记忆标记物， 标记物，用于CFSE研究，以及用于分选从淋巴和淋巴细胞过继转移的T细胞亚群， GVHD小鼠和延迟DLI受体的非淋巴组织。项目2（杨）将调查 IFN-γ将GVL效应与供体CDS T细胞的GVHD诱导活性分开的机制 在同种异体HCT接受者中。该计划将需要一个细胞分选机约2天/月，以纯化供体， 受体小鼠细胞，从大约30只小鼠/年获得，用于移植。对于项目3（萨克斯）， 核心A将用于供体DLI接种物的特异性T细胞亚群消耗和DLI接种物的分离。 HCT后，每月从嵌合受体的PBMC中采集供体型和宿主型T细胞 调节细胞的体外和体内机制研究的目的。对于项目4（Scadden），结果 将监测用G-CSF动员治疗的患者中PTH给药对CD 34+细胞产量的影响。 还将监测接受PTH治疗的移植后患者的免疫重建。为 与项目4核心A相关的小鼠研究将根据T细胞 分化抗原和MHC特异性。大约需要20种和45种分析 项目

Core A contains a Becton-Dickinson DiVa cell sorter and LSR analyzer. The DiVa is a 13-parameter 4-way 5-laser cell sorter. The LSR has excitation lines in the ultraviolet, visible and red spectra. The Core Leader was recently awarded (1/1/05) a SIG to build a 6-laser, 20-detector LSR-2 which will be available for this program. For Project 1 (Sykes), Core A will be used for 9-color flow cytometry studies required to assess activation and memory markers on GVH-reactive and non-reactive T cell subsets identified with specific markers, for CFSE studies, and for sorting of T cell subsets to be adoptively transferred from lymphoid and non-lymphoid tissues of GVHD mice and delayed DLI recipients. Project 2 (Yang) will investigate the mechanisms by which IFN-gamma separates GVL effects from GVHD-inducing activity of donor CDS T cells in allogeneic HCT recipients. The program will need a cell sorter about 2 days/month to purify donor from recipient mouse cells, obtained from approximately 30 mice/yr, for transplantation. For Project 3 (Sachs), Core A will be used for the specific T-cell subset depletion of the donor DLI inoculum and the separation of donor-type and host-type T-cells from PBMC of chimeric recipients at monthly intervals, following HCT for the purpose of in vitro and in vivo mechanistic studies of regulatory cells. For Project 4 (Scadden), the result of PTH administration on CD34+ cell yields in patients treated with G-CSF mobilization will be monitored. The immune reconstitution of post-transplant patients treated with PTH will also be monitored. For the murine studies related to Project 4, Core A will assess and separate lymphocytes based upon T-cell differentiation antigens and MHC specificity. Approximately 20 sorts and 45 analyses will be needed for this project.