In vivo imaging of T Cells using engineered antibodies and PET

使用工程抗体和 PET 对 T 细胞进行体内成像

• 批准号: 8786848
• 负责人: Anna M Wu
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8786848
• 关键词: Address; Affinity; Agonist; Antibodies; Antigens; Autoimmunity; B-Lymphocytes; Binding; Biodistribution; Biological Markers; Blood; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cardiovascular Diseases; Cell Therapy; Cell surface; Cells; Characteristics; Chimeric Proteins; Clinic; Cysteine; Cytosine; Cytotoxic T-Lymphocytes; Detection; Development; Diabetes Mellitus; Disease; Disease model; Disulfides; Dose; Engineering; Epitopes; Exhibits; Flow Cytometry; Future; Generations; Goals; Health; Helper-Inducer T-Lymphocyte; Human; Hybridomas; Image; Imagery; Immune; Immune response; Immune system; Immunoglobulin Fragments; Immunohistochemistry; Immunotherapeutic agent; Immunotherapy; Infiltration; Inflammation; Injection of therapeutic agent; Kinetics; Label; Lead; Length; Life; Location; Lymphoid; Lymphoid Tissue; MS4A1 gene; Malignant Neoplasms; Modeling; Monitor; Monoclonal Antibodies; Mus; Oligonucleotides; Organ; Organism; Phenotype; Play; Positron; Positron-Emission Tomography; Pre-Clinical Model; Process; Production; Proliferating; Protein Engineering; Proteins; Protocols documentation; Radioimmunoconjugate; Radiolabeled; Rattus; Resolution; Role; Spleen; Staging; T-Lymphocyte; Therapeutic; Therapy Evaluation; Time; Tissues; Tracer; Wild Type Mouse; Work; antibody engineering; base; clinical application; dimer; drug development; effective therapy; fluorodeoxyglucose; imaging modality; immune activation; immune function; immunoreactivity; improved; in vivo; in vivo imaging; interest; lymph nodes; macrophage; molecular imaging; mouse model; neutrophil; novel; pre-clinical; public health relevance; radiotracer; research study; small molecule; success; tool; tumor

DESCRIPTION (provided by applicant): Inflammation and immune responses play a central role in most of the major diseases that impact human health, including cancer, cardiovascular disease, autoimmunity, neuroinflammatory processes, diabetes and other diseases. In addition, immunomodulatory therapies and adoptive cell therapies have become established as powerful therapeutic approaches in cancer. The ability monitor and distinguish immune cell subsets (e.g. T and B lymphocytes, macrophages, neutrophils, etc.) noninvasively in vivo will be invaluable to the development of effective treatments. Positron emission tomography (PET) provides highly sensitive, non- invasive and quantitative imaging, which can be combined with antibody-based probes to provide molecular imaging of tissues and cells in vivo based on cell surface phenotype. Furthermore, engineered antibody fragments (minibodies and cys-diabodies) have been developed with optimal kinetics enabling rapid in vivo targeting and clearance for imaging applications by immunoPET. In the proposed work, immunoPET will be developed to address the challenge of direct imaging of immune cell subsets. Engineered antibody fragments will be optimized for in vivo detection, imaging, and quantitation of T lymphocytes that express CD8, a marker that is a hallmark of cytotoxic T cells. Specific Aim 1 will focus on production of stable, optimized anti-CD8 minibodies and cys-diabodies derived from rat anti-mouse monoclonal antibodies. Specific Aim 2 will establish the optimal imaging agent, dose, and imaging protocol for visualization of the lymphoid organs in normal (wild-type) mice. Specific Aim 3 will focus on quantitation of T cells in normal and immune-stimulated mice to determine the parameters and limits of detection in mouse models. Once validated, immunoPET probes for CD8 T cells will be valuable tools to study immune processes in mouse models, providing a deeper understanding of immune responses in living organisms, and will open the door to broad methods for imaging and quantitating immune cell subsets in preclinical models and eventually humans.

描述（由申请人提供）：炎症和免疫反应在影响人类健康的大多数主要疾病中发挥核心作用，包括癌症、心血管疾病、自身免疫、神经炎症过程、糖尿病和其他疾病。此外，免疫调节疗法和过继细胞疗法已成为癌症强有力的治疗方法。在体内无创监测和区分免疫细胞亚群（如T淋巴细胞、B淋巴细胞、巨噬细胞、中性粒细胞等）的能力对开发有效的治疗方法将是非常宝贵的。正电子发射断层扫描（PET）提供高灵敏度、非侵入性和定量成像，可与基于抗体的探针结合，提供基于细胞表面表型的体内组织和细胞的分子成像。此外，工程抗体片段（微型体和半胱氨酸-糖尿病体）已经开发出最佳的动力学，能够快速在体内靶向和清除，用于免疫pet成像应用。在提出的工作中，免疫pet将被开发来解决免疫细胞亚群直接成像的挑战。工程化抗体片段将被优化用于体内检测、成像和定量表达CD8的T淋巴细胞，CD8是细胞毒性T细胞的标志。特异性目标1将专注于从大鼠抗小鼠单克隆抗体中提取稳定的、优化的抗cd8微体和半胱氨酸糖体。特异性目标2将建立正常（野生型）小鼠淋巴器官可视化的最佳显像剂、剂量和成像方案。特异性目标3将集中于正常和免疫刺激小鼠的T细胞定量，以确定小鼠模型的参数和检测限。一旦验证，CD8 T细胞的免疫pet探针将成为研究小鼠模型免疫过程的有价值的工具，提供对生物体免疫反应的更深入了解，并将为临床前模型和最终人类的免疫细胞亚群成像和定量的广泛方法打开大门。