Cell Processing and Immune Assessment

细胞处理和免疫评估

• 批准号: 8462451
• 负责人: JEROME RITZ
• 金额: $ 28.29万
• 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/8462451
• 关键词: Antigen Targeting; Antigens; Aspirate substance; Autologous; Autologous Stem Cell Transplantation; Biological Assay; Biopsy; Blood Cells; Bone Marrow; Bone Marrow Cells; CA-15-3 Antigen; Cell Communication; Cell physiology; Cells; Clinical; Clinical Trials; Core Facility; Cytomegalovirus; Development; Dissection; Elements; Enrollment; Enzyme-Linked Immunosorbent Assay; Excision; Flow Cytometry; Fluorochrome; Goals; Growth; Human Herpesvirus 4; Immune; Immune response; Immune system; Immunity; Immunologic Monitoring; Immunotherapy; Individual; Informed Consent; Infusion procedures; Innovative Therapy; Laboratories; Lead; Measurement; Measures; Methods; Monoclonal Antibodies; Multiple Myeloma; Natural Immunity; Outcome; Patients; Peptides; Plasma; Population; Procedures; Production; Research Project Grants; Resources; Sampling; Serum; T-Cell Receptor; T-Lymphocyte; Therapeutic; Tumor Antigens; Vaccines; adaptive immunity; base; conventional therapy; cytokine; enzyme linked immunospot assay; immune function; in vivo; neoplastic cell; programs; reconstitution; research study; response; tumor microenvironment

A central theme of the research projects in this program is that the growth of myeloma tumor cells is modulated through multiple and functionally diverse interactions with the tumor cell microenvironment. Many of these interactions involve elements of innate and adaptive immunity that can either promote or inhibit the growth of myeloma cells in vivo. A better understanding of both negative and positive immune system interactions can lead to the development of innovative therapies for patients with myeloma. This is a goal of each of the 3 research projects and this core will support each project by providing a centralized laboratory resource for immune monitoring and manufacturing immune cells for adoptive cellular therapy. For patients with myeloma who receive conventional therapies as well as those enrolled on specific clinical trials supported by this program, we have established procedures for obtaining samples of myeloma tumor cells and normal immune cells for research studies (described in Core A). Bone marrow aspirates and biopsies and well as peripheral blood cells are obtained after informed consent and viable cells, plasma and serum are cryopreserved at regular intervals. Using these samples Core C will provide a detailed analysis of immune function using a variety of methods that provide a quantitative assessment of specific populations as well as their level of maturation and function. Quantitative analysis of circulating immune cells as well as bone marrow cells and myeloma cells is determined by multi-parameter flow cytometry with a panel of fluorochrome-conjugated monoclonal antibodies. Cytokine production by distinct subsets of immune cells is also determined by flow cytometry. Levels of individual cytokines are measured by bead-based multiplex assays or by ELISA. Reconstitution of T cell receptor repertoire is examined by TCR VB spectratyping. Thymic function is evaluated by quantitative PCR for T-cell receptor excision circles (TREC). T cell immunity to specific target antigens such as CMV and EBV or to myeloma-associated tumor antigens such as MUC-1 is determined by ELISPOT or with fluorescent-HLA-peptide-conjugates. Results of these assays are correlated with other parameters of immune function as well as with clinical outcomes. For patients enrolled on the clinical trials of myeloma/DC fusion vaccine and T cell infusions described in Project 3, this core will have the additional responsibility for clinical scale manufacturing of large numbers of autologous T cells for adoptive T cell infusions. The specific aims of this core are listed below: 1. To provide phenotypic and functional measurements of immune function and response to immune therapies. 2. Manufacture patient T cells for adoptive immune therapy

该计划中研究项目的一个核心主题是，骨髓瘤肿瘤细胞的生长是通过与肿瘤细胞微环境的多种且功能上不同的相互作用来调节的。这些相互作用中的许多涉及先天和适应性免疫的元素，这些元素可以促进或抑制体内骨髓瘤细胞的生长。更好地了解负面和阳性免疫系统相互作用可以导致骨髓瘤患者的创新疗法发展。这是三个研究项目中的每个项目的一个目标，该核心将通过为免疫监测和制造免疫细胞提供用于过养的细胞治疗的集中式实验室资源来支持每个项目。 对于接受常规疗法的骨髓瘤患者以及该程序支持的特定临床试验的患者，我们已经建立了用于获取骨髓瘤肿瘤细胞和正常免疫细胞样品进行研究的程序（核心A中描述）。在知情同意书和生存的细胞，血浆和血清后，骨髓骨髓抽吸和活检以及以及周围血细胞是定期冷冻保存的。使用这些样品核心C将使用各种方法对特定人群进行定量评估的各种方法提供详细的免疫功能分析 以及它们的成熟和功能水平。通过多参数流式细胞仪，用一组荧光偶联的单克隆抗体来确定循环免疫细胞以及骨髓细胞和骨髓瘤细胞的定量分析。通过流式细胞术决定了由免疫细胞的不同子集的细胞因子产生。单个细胞因子的水平通过基于珠的多重测定或ELISA测量。通过TCR VB频谱检查TCR受体库的重建。通过定量PCR评估胸腺功能的T细胞受体切除圈（TREC）。 T细胞对特定靶抗原（例如CMV和EBV）或对骨髓瘤相关的肿瘤抗原（如MUC-1）的免疫性是由ELISPOT或荧光HLA肽偶联物确定的。这些测定的结果与免疫功能的其他参数以及临床结果相关。对于参加项目3中描述的骨髓瘤/DC融合疫苗和T细胞输注的临床试验的患者，该核心将对大量自体T细胞的临床规模产生额外的责任。该核心的具体目的如下： 1。提供免疫功能的表型和功能测量以及对免疫疗法的反应。 2。生产患者T细胞进行过养免疫治疗