Biobanking and Immunologic Monitoring

核心2项目摘要/摘要 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(HCT)和随后的免疫干预预防或治疗慢性 GVHD。移植前，外周血单个核细胞(PBMC)、骨髓和血清或血浆 是从参加本计划项目支持的临床试验的所有患者获得的。PBMC、血浆和 血清也从接受常规异基因干细胞移植和正常干细胞移植的患者中获得。 这些人的细胞捐赠者。在红细胞压积试验后，采集外周血、血浆和DNA样本 有规律的间隔。所有这些样品都经过处理，冷冻保存，输入计算机化的库存 提供给这个项目中的所有调查人员。HCT组织库库存也非常大 曾在Dana-Farber接受过异基因红细胞移植的患者的连续样本数量 癌症研究所、布里格姆妇女医院和波士顿儿童医院。关于这些项目的临床结果数据 患者可在我们的HCT数据库中找到。此带注释的样本清单也可供计划使用 调查人员。最后，组织库清单包括从以前的临床试验中获得的系列样本。 评估小剂量IL-2和其他新的治疗药物对慢性移植物抗宿主病患者的影响 (CGVHD)。所有储存的样本均来自已提供样本知情同意书的病人。 为研究收集资料。监测异基因血细胞移植后的免疫重建及干预 为了预防或治疗移植物抗宿主病，这个核心使用了多参数流式细胞术和一组荧光色素- 结合的单抗或与金属结合的抗体组合的质量细胞术。如果需要， 已知在免疫重建中起重要作用的细胞因子的血浆浓度通过以下方法测量 可以通过Luminex检测到酶联免疫吸附试验或细胞因子组合。针对特定靶点的T细胞免疫重建 CMV和EBV等抗原由ELISPOT和HLA四聚体确定。每种化验方法的结果 与免疫功能的其他参数以及临床结果相关。

Core 2 Project Summary/Abstract 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) and following immune interventions to prevent or treat chronic GVHD. 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 patients undergoing routine allogenic stem cell transplantation and 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. The HCT tissue bank inventory also has a very large number of serial samples from patients who have previously undergone allogeneic HCT at the Dana-Farber Cancer Institute, Brigham and Women’s Hospital and Boston Children’s Hospital. Clinical outcome data on these patients is available in our HCT database. This inventory of annotated samples is also available to Program investigators. Finally, the tissue bank inventory includes serial samples obtained from previous clinical trials evaluating low dose IL-2 and other novel therapeutic agents in patients with chronic graft versus host disease (cGVHD). All banked samples have been collected from patients who have provided informed consent for sample collection for research studies. To monitor immune reconstitution after allogeneic HCT as well as interventions to prevent or treat GVHD, this core utilizes multi-parameter flow cytometry with a panel of fluorochrome- conjugated monoclonal antibodies or mass cytometry with panels of metal-conjugated antibodies. If needed, plasma concentrations of cytokines known to play important roles in immune reconstitution are measured by ELISA or panels of cytokines can be measured by Luminex. Reconstitution of T cell immunity to specific target antigens such as CMV and EBV is determined by ELISPOT and HLA-tetramers. Results of each of these assays are correlated with other parameters of immune function as well as with clinical outcomes.