Core B: Cell Banking and Immune Assessment Core

项目摘要/摘要核心B 这个核心的目的是提供一个集中的实验室资源，用于获取、低温保存和 分发患者样本并监测异基因移植后免疫重建的动力学 造血干细胞移植。移植前，外周血单核细胞 (PBMC)、骨髓和血清或血浆是从所有参加支持的临床试验的患者那里获得的 由本计划项目提供。PBMC、血浆和血清也是从正常干细胞捐赠者那里获得的 个人。在HCT后，定期采集外周血、血浆和DNA样本。所有的 这些样品经过处理、冷冻保存，输入计算机化的库存，并向所有人提供 这个项目中的调查人员。支持项目1中的临床试验，免疫学的影响 通过对定义变化的特定淋巴细胞群体的系列评估来评估干预措施 在数量上以及它们的激活程度、成熟程度等功能特征上。表型 循环淋巴细胞的分析主要利用多参数流式细胞术 荧光素偶联的单抗。已知的血浆细胞因子浓度在其中起重要作用 用双抗体夹心法检测BAFF、IL-2在免疫重建中的作用。B细胞和T细胞受体的重建 通过对免疫球蛋白和TCRV基因进行深入测序来检查曲目。胸腺功能是 用定量聚合酶链式反应测定T细胞受体切除环(TREC)进行评价。T细胞的重建 对CMV和EBV等特定靶抗原的免疫力由ELISPOT和HLA四聚体决定。在……里面 项目3，患者血浆用于识别针对HY抗原或其他同种或自身抗原的抗体。 每项检测的结果都与免疫功能的其他参数以及 临床结果。

Project Summary/Abstract Core B The purpose of this core is to provide a centralized laboratory resource for obtaining, cryopreserving and distributing patient samples and to monitor the kinetics of immune reconstitution following allogeneic hematopoietic stem cell transplantation (HCT). Prior to transplantation, peripheral blood mononuclear cells (PBMC), bone marrow and serum or plasma are obtained from all patients enrolled on clinical trials supported by this Program Project. PBMC, plasma and serum are also obtained from normal stem cell donors for these individuals. Following HCT, peripheral blood, plasma and DNA samples are obtained at regular intervals. All of these samples are processed, cryopreserved, entered into a computerized inventory and made available to all of the investigators in this program. To support clinical trials in Project 1, the effects of immunologic interventions are evaluated through serial assessment of specific lymphocyte populations that define changes in number as well as their level of activation, maturation and other functional characteristics. Phenotypic analysis of circulating lymphocytes primarily utilizes multi-parameter flow cytometry with a panel of fluorochrome-conjugated monoclonal antibodies. Plasma concentrations of cytokines known to play important roles in immune reconstitution (BAFF, IL-2) are measured by ELISA. Reconstitution of B and T cell receptor repertoire is examined by in depth sequencing of immunoglobulin and TCRV genes. Thymic function is evaluated by quantitative PCR measurement of T-cell receptor excision circles (TREC). Reconstitution of T cell immunity to specific target antigens such as CMV and EBV is determined by ELISPOT and HLA-tetramers. In Project 3, patient plasma is used to identify antibodies specific for HY antigens or other allo or auto antigens. Results of each of these assays are correlated with other parameters of immune function as well as with clinical outcomes.