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着核心作用，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的强大治疗方法。在体内无创地监测和区分免疫细胞亚群（例如 T 和 B 淋巴细胞、巨噬细胞、中性粒细胞等）的能力对于开发有效的治疗方法具有不可估量的价值。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，可以与基于抗体的探针结合，根据细胞表面表型提供体内组织和细胞的分子成像。此外，我们还开发了具有最佳动力学的工程化抗体片段（微型抗体和 cys-双抗体），可实现免疫 PET 成像应用中的快速体内靶向和清除。在拟议的工作中，将开发免疫PET来解决免疫细胞亚群直接成像的挑战。工程抗体片段将针对表达 CD8 的 T 淋巴细胞的体内检测、成像和定量进行优化，CD8 是细胞毒性 T 细胞的标志物。具体目标 1 将重点关注源自大鼠抗小鼠单克隆抗体的稳定、优化的抗 CD8 微型抗体和 cys-双抗体的生产。具体目标 2 将建立最佳显像剂、剂量和成像方案，用于正常（野生型）小鼠淋巴器官的可视化。具体目标 3 将重点关注正常小鼠和免疫刺激小鼠中 T 细胞的定量，以确定小鼠模型中的参数和检测限。一旦经过验证，CD8 T 细胞的免疫 PET 探针将成为研究小鼠模型免疫过程的宝贵工具，提供对生物体免疫反应的更深入了解，并将为临床前模型乃至人类免疫细胞亚群成像和定量的广泛方法打开大门。