Novel B Cell Reagents

新型 B 细胞试剂

• 批准号: 8681324
• 负责人: Max Dale Cooper
• 金额: $ 33.67万
• 依托单位: EMORY UNIVERSITY
• 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/8681324
• 关键词: Affinity; Antibodies; Antibody Formation; Antibody Repertoire; Antibody Specificity; Antigen Targeting; Antigens; Architecture; B-Lymphocytes; Biological Markers; Blast Cell; Blood; Blood Circulation; CD4 Positive T Lymphocytes; Cell Line; Cells; Cellular biology; Characteristics; Chimeric Proteins; Control Animal; Evaluation; Expression Library; Extracellular Domain; Fc Receptor; Fc domain; Flow Cytometry; Generations; Goals; HIV; HIV Envelope Protein gp120; Helper-Inducer T-Lymphocyte; Human; Humoral Immunities; Immune; Immune response; Immune system; Immunoglobulin G; Immunoglobulins; Individual; Infection; Instruction; Invaded; Lampreys; Larva; Lead; Leucine-Rich Repeat; Lymphocyte; Lymphoid Tissue; Macaca; Macaca mulatta; Maintenance; Memory; Memory B-Lymphocyte; Monoclonal Antibodies; Mucous Membrane; Mus; Pathogenesis; Petromyzon marinus; Phenotype; Plasma; Plasma Cells; Plasmablast; Population; Primates; Proteins; Reagent; Recombinants; Research; Research Project Grants; Role; SIV; SIV Vaccines; Specificity; Structure; Surface; Tissues; Vaccinated; Vaccination; Vaccines; Vertebrates; Virus; base; exhaust; improved; neutralizing antibody; neutrophil; novel; pathogen; research study; response; vaccination strategy; vaccine evaluation; virology

Memory B cells are key contributors to the indiction and maintenance of humoral immunity. Characterization of the memory B cells response to SIV will be important in an vaccine strategy to elicit a neutralizing antibody response. In this research application we propose to develop reagents to specifically identify macaque memory B cell populations. In proof of principle studies we have produced monoclonal lamprey VLR antibodies that are specific for memory B cells in humans. We propose to develop novel monoclonal VLR antibodies to macaque memory B cells. Unlike mammalian antibodies, the VLR antibodies of the jawless sea lampreys utilize leucine-rich repeats as basic structural units. The radically different protein architecture combined with the large evolutionary distance ofthe sea lamprey allows VLR antibodies to be made that recognize antigenic structures which are not detetcted by mammalian immunoglobulin-based antibodies for structural or tolerogenic reasons. To generate monoclonal VLR antibodies, we will immunize sea lamprey larvae with purified primary B cells or B cell lines. VLR antibody expression libraries will be screened by flow cytometry and monoclonal VLR antibodies that specifically interact with memory B cells or subpopulations thereof will be used as affinity reagents to identify the antigen recognized by the VLR antibody. The VLR antibodies from this core will be provided to research groups to analyze the memory B cell responses to vaccination or SIV challenge. In a complementary approach, we will generate conventional mammalian monoclonal antibodies to FCRL4, an immunoreceptor that In humans is expressed by a subpopulation of mucosa associated memory B cells with enrichment in antibody specificity for the HIV gp120 protein. A fusion protein consisting ofthe extracellular domain of FCRL4 fused to the Fc-domain of IgG will serve as the immunogen and the specific monoclonal antibodies will be identified by flow cytometry. The long term goal of these studies is to generate novel reseach reagents that will facilitate the analysis of memory B cells responding to SIV challenge or vaccination.

记忆 B 细胞是体液免疫的指示和维持的关键贡献者。表征 记忆 B 细胞对 SIV 的反应对于诱发中和抗体的疫苗策略非常重要 回复。在这项研究应用中，我们建议开发试剂来特异性识别猕猴 记忆 B 细胞群。在原理研究证明中，我们生产了单克隆七鳃鳗 VLR 专门针对人类记忆 B 细胞的抗体。我们建议开发新型单克隆VLR 猕猴记忆 B 细胞抗体。与哺乳动物抗体不同，无颌海的 VLR 抗体 七鳃鳗利用富含亮氨酸的重复序列作为基本结构单元。截然不同的蛋白质结构 结合海七鳃鳗的长进化距离，可以制造出 VLR 抗体， 识别哺乳动物免疫球蛋白抗体无法检测到的抗原结构 结构或耐受性原因。为了产生单克隆 VLR 抗体，我们将免疫海七鳃鳗 具有纯化的原代 B 细胞或 B 细胞系的幼虫。流式筛选VLR抗体表达文库 与记忆 B 细胞或亚群特异性相互作用的细胞计数法和单克隆 VLR 抗体 其将用作亲和试剂来鉴定VLR抗体所识别的抗原。访问寄存器 来自该核心的抗体将提供给研究小组，以分析记忆 B 细胞对 疫苗接种或 SIV 攻击。通过补充方法，我们将产生传统的哺乳动物 FCRL4 的单克隆抗体，FCRL4 是一种在人类中由以下亚群表达的免疫受体 粘膜相关记忆 B 细胞，富含 HIV gp120 蛋白的抗体特异性。一个 由 FCRL4 的胞外结构域与 IgG 的 Fc 结构域融合组成的融合蛋白将作为 免疫原和特异性单克隆抗体将通过流式细胞术鉴定。长期来看 这些研究的目标是产生新的研究试剂，以促进记忆 B 细胞的分析 应对 SIV 攻击或疫苗接种。