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的单细胞技术，以分析恒河猴对SIV疫苗接种或感染的持续免疫应答期间的特异性浆母细胞。我们将产生详细的信息，动力学，免疫球蛋白同种型useage和plasmablast的反应诱导在全身水平以及在粘膜部位的表型，并确定这些反应是如何调制使用不同的佐剂。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，鉴定浆母细胞靶向的特异性表位，并确定单个抗体对其抗原的亲和力/亲合力。此外，我们将描述分离抗体的功能特性，包括中和作用、中和宽度以及非中和性、抗体介导的细胞溶解机制。此外，特别令人感兴趣的单克隆抗体，即广泛中和的，也可能是未来在恒河猴中进行被动转移实验的优秀候选物，并且还对人类HIV的治疗和疫苗设计具有影响。