Interrogating the Plasmablast Response

询问浆母细胞反应

• 批准号: 8883357
• 负责人: Jens Peter Wrammert
• 金额: $ 32.5万
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8883357
• 关键词: Address; Adjuvant; Affinity; Animals; Antibodies; Antibody Response; Antigens; Avidity; B-Lymphocytes; Biological Assay; Blood; Blood specimen; Bone Marrow; Cell Separation; Cells; Cellular biology; Characteristics; Collaborations; Development; Epitopes; Future; Generations; Genes; Goals; HIV; HIV Infections; HIV vaccine; Homing; Human; Immune; Immune response; Immunoglobulin A; Immunoglobulin G; Immunoglobulin Isotypes; Immunoglobulin M; Immunoglobulin Somatic Hypermutation; Immunoglobulins; Individual; Infection; Instruction; Kinetics; Laboratories; Light; Macaca; Macaca mulatta; Mediating; Methods; Modeling; Monoclonal Antibodies; Mucous Membrane; Phenotype; Plasmablast; Property; Reagent; Recombinants; Regimen; Research Design; Reverse Transcriptase Polymerase Chain Reaction; SIV; SIV Vaccines; Sampling; Sorting - Cell Movement; Specificity; Spleen; System; Technology; Therapeutic; Time; Vaccination; Vaccine Design; Vaccines; Vagina; Viral; Virus; direct application; env Gene Products; enzyme linked immunospot assay; gag Gene Products; human monoclonal antibodies; interest; mucosal site; neutralizing antibody; neutralizing monoclonal antibodies; new technology; novel; programs; prophylactic; prototype; rectal; research study; response; screening; single cell analysis; therapy design; vaccine trial; virology

The goal of this Core is to establish novel single cell technology to analyze specific plasmablasts during an ongoing immune response to SIV vaccination or infection in rhesus macaques. We will generate detailed information about the dynamics, immunoglobulin isotype useage and phenotype of the plasmablast responses induced both at a systemic level as well as at mucosal sites, and determine how these responses are modulated by the use of different adjuvants. The single cell analysis will allow us to interrogate the immunoglobulin repertoire of the ongoing response, identify the specific epitopes targeted by the plasmablasts and define the affinity/avidity ofthe individual antibodies against their antigen. Furthermore we will characterize the functional properties ofthe isolated antibodies with regards to neutralization, breadth of neutralization as well as non-neutralizing, antibody mediated cytolytic mechanisms. Furthermore, particularly interesting monoclonal antibodies, i.e. broadly neutralizing, might also be excellent candidates for future passive transfer experiments in rhesus macaques and also have implications for therapy and vaccine design for HIV in humans.

该核心的目标是建立新型单细胞技术，以分析恒河猴对 SIV 疫苗接种或感染的持续免疫反应期间的特定浆母细胞。我们将生成有关在全身水平以及粘膜部位诱导的浆母细胞反应的动力学、免疫球蛋白同种型使用和表型的详细信息，并确定如何通过使用不同的佐剂来调节这些反应。单细胞分析将使我们能够询问正在进行的反应的免疫球蛋白库，识别浆母细胞靶向的特定表位，并确定单个抗体针对其抗原的亲和力/亲合力。此外，我们将表征分离的抗体在中和、中和广度以及非中和、抗体介导的细胞溶解机制方面的功能特性。此外，特别有趣的单克隆抗体，即广泛中和，也可能是未来在恒河猴中进行被动转移实验的优秀候选者，并且对人类艾滋病毒的治疗和疫苗设计也具有影响。