Cell Banking and Immune Assessment

细胞库和免疫评估

• 批准号: 8257937
• 负责人: JEROME RITZ
• 金额: $ 34.76万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8257937
• 关键词: Allogenic; Antigen Targeting; Biological Assay; Biopsy; Blood specimen; Bone Marrow; Cells; Clinical; Clinical Trials; Confidentiality of Patient Information; Cryopreservation; Cytomegalovirus; DNA; Engraftment; Enrollment; Equipment and supply inventories; Excision; Flow Cytometry; Fluorochrome; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Immune; Immunity; Immunofluorescence Immunologic; Immunologics; Individual; Informed Consent; Kinetics; Laboratories; Lymphocyte; Measurement; Methods; Monitor; Monoclonal Antibodies; Mononuclear; Outcome; Patients; Peripheral Blood Mononuclear Cell; Plasma; Population; Population Control; RNA; Research Personnel; Resources; Sampling; Serum; Skin; Stem cell transplant; Stem cells; T-Cell Receptor; T-Lymphocyte; Time; Toxic effect; Transplantation; Whole Blood; cell bank; computerized; effective therapy; enzyme linked immunospot assay; graft vs host disease; immune function; patient privacy; peripheral blood; programs; reconstitution; research study

The purpose of this core is to provide a centralized laboratory resource for cryopreservation and distribution of patient samples and to monitor the kinetics of immunologic reconstitution following allogeneic hematopoietic stem cell transplantation (HSCT). 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 and plasma or serum are also obtained from normal stem cell donors for these individuals. Following HSCT, PBMC, DNA and plasma samples are cryopreserved at regular intervals. These samples are made available to all of the investigators in this program. Immune reconstitution after HSCT is evaluated through a variety of methods that provide a quantitative assessment of specific lymphocyte populations as well as their level of maturation and function. Quantitative analysis of circulating lymphocytes is determined by multi-parameter flow cytometry with a panel of fluorochrome-conjugated monoclonal antibodies. This is primarily evaluated using fresh samples and semi-automated methods for immunofluorescence analysis of whole blood. Assays of immune function primarily utilize cryopreserved PBMC. Reconstitution of T cell receptor repertoire is examined by the method of TCR Vp spectratyping. Thymic function is evaluated by quantitative PCR measurement of T-cell receptor excision circles (TREC) in PBMC. Reconstitution of T cell immunity to specific target antigens such as CMV and EBV is determined by ELISPOT. Results of each of these assays can be correlated with other parameters of immune function as well as with clinical outcomes. The availability of pre-HSCT samples as well as normal donor samples provides additional control populations and will allow additional comparisons. This core has 3 Specific Aims: 1. To obtain, isolate and cryopreserve mononuclear cells, DNA, RNA, plasma and serum components from patient samples for subsequent study by program investigators. 2. To maintain a computerized inventory of banked samples, assure patient privacy and distribute samples in a timely and equitable fashion to laboratory investigators for experiments proposed in this application. 3. To provide phenotypic and functional measurements of immune reconstitution in peripheral blood, bone marrow and targeted skin biopsies following allogeneic HSCT.

该核心的目的是为患者样本的冷冻保存和分配提供集中的实验室资源，并监测同种异体造血干细胞移植（HSCT）后免疫重建的动力学。移植前，从参加本计划支持的临床试验的所有患者中获取外周血单核细胞 (PBMC)、骨髓和血清或血浆。 PBMC和血浆或血清也取自正常干细胞 这些人的捐助者。 HSCT 后，PBMC、DNA 和血浆样本会定期冷冻保存。这些样本可供该计划中的所有研究人员使用。 HSCT 后的免疫重建可通过多种方法进行评估，这些方法可对特定淋巴细胞群及其成熟和功能水平进行定量评估。使用一组荧光染料偶联的单克隆抗体通过多参数流式细胞术对循环淋巴细胞进行定量分析。这主要是使用新鲜样本和全血免疫荧光分析的半自动方法进行评估。免疫功能测定主要利用冷冻保存的 PBMC。通过 TCR Vp 谱型分析方法检查 T 细胞受体库的重建。通过定量 PCR 测量 PBMC 中的 T 细胞受体切除环 (TREC) 来评估胸腺功能。 T 细胞对特定靶抗原（如 CMV 和 EBV）免疫的重建由 ELISPOT 确定。每项检测的结果都可以与免疫功能的其他参数以及临床结果相关。 HSCT 前样本以及正常供体样本的可用性提供了额外的对照人群，并将允许进行额外的比较。 该核心有 3 个具体目标： 1. 从患者样本中获取、分离和冷冻保存单核细胞、DNA、RNA、血浆和血清成分，供项目研究人员进行后续研究。 2. 为了维护存储样本的计算机化库存，确保患者隐私并及时且公平地将样本分发给实验室研究人员以进行本申请中提出的实验。 3. 提供同种异体 HSCT 后外周血、骨髓和靶向皮肤活检中免疫重建的表型和功能测量。