Functional Antibody Study

功能性抗体研究

• 批准号: 10588239
• 负责人: Alison K Criss
• 金额: $ 22.63万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-26 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10588239
• 关键词: Adjuvant; AlamarBlue; Antibodies; Antibody Response; Antibody titer measurement; Antigens; Bacteria; Bacterial Antibodies; Binding; Biological Assay; Biology; Cells; Complement; Dyes; Ensure; Evaluation; Flow Cytometry; Goals; Gonorrhea; HL60; Human; Image; Immune Sera; Immune response; Immunity; Immunize; Immunoglobulin G; Immunoglobulin M; Infection; Laboratories; Licensure; Measures; Mediating; Metabolic; Mus; Neisseria; Neisseria gonorrhoeae; Neisseria meningitidis; Opsonin; Pathogenicity; Phagocytes; Progranulocytes; Public Health; Research; Research Project Grants; Resolution; Role; Saline; Serogroup B Neisseria meningitidis; Serum; Services; Source; Surface; Universities; Vaccination; Vaccine Antigen; Vaccinee; Vaccines; Virginia; Virus-like particle; Work; antibody test; bactericide; global health; gonorrhea vaccine; health goals; interest; novel vaccines; response; success; vaccine development; Adjuvant; AlamarBlue; Antibodies; Antibody Response; Antibody titer measurement; Antigens; Bacteria; Bacterial Antibodies; Binding; Biological Assay; Biology; Cells; Complement; Dyes; Ensure; Evaluation; Flow Cytometry; Goals; Gonorrhea; HL60; Human; Image; Immune Sera; Immune response; Immunity; Immunize; Immunoglobulin G; Immunoglobulin M; Infection; Laboratories; Licensure; Measures; Mediating; Metabolic; Mus; Neisseria; Neisseria gonorrhoeae; Neisseria meningitidis; Opsonin; Pathogenicity; Phagocytes; Progranulocytes; Public Health; Research; Research Project Grants; Resolution; Role; Saline; Serogroup B Neisseria meningitidis; Serum; Services; Source; Surface; Universities; Vaccination; Vaccine Antigen; Vaccinee; Vaccines; Virginia; Virus-like particle; Work; antibody test; bactericide; global health; gonorrhea vaccine; health goals; interest; novel vaccines; response; success; vaccine development

PROJECT SUMMARY/ABSTRACT The overall objective of Core D: Functional Antibody Study Core is to provide a consistent, experimentally controlled and validated platform for evaluating functional antibody responses after vaccination with Neisseria gonorrhoeae antigens that are identified through the projects of the Gonorrhea Vaccine Cooperative Research Center (GV CRC). Serum bactericidal activity and opsonophagocytosis have been implicated in the protective immune response against pathogenic Neisseria, and both depend upon antibody binding to the bacterial surface. Evaluation of these three parameters of the functional antibody response is critical to understanding if a vaccine has the potential to elicit protective immunity. Therefore, Core D is critical to the success of the GV CRC. Core D will provide services to all four research projects and will interact routinely with the other Cores of this CRC, which offer distinct yet complementary expertises. For Core D to enable successful completion of the projects in the GV CRC, we propose three Specific Aims: 1) Bacterial Antibody Surface Binding: We will use imaging flow cytometry to quantify the ability of antibodies in sera from immunized mice to bind to the surface of intact N. gonorrhoeae. Bacteria of diverse strain backgrounds will be evaluated. 2) Serum Bactericidal Activity (SBA): We will measure the ability of antibodies in sera from immunized mice or humans immunized with N. meningitidis serogroup B vaccine to elicit SBA against a panel of N. gonorrhoeae strains, using antibody-depleted pooled normal human serum as the source of active complement. In addition to conventional colony count, we will pilot and optimize a high-throughput quantitative assay using a fluorometric metabolic dye as a surrogate measure of SBA. 3) Opsonphagocytic Activity (OPA): We will measure the ability of the antibodies in sera from immunized mice and N. meningitidis serogroup B-immunized humans to enhance OPA-dependent killing of N. gonorrhoeae, using HL-60 human promyelocytes as the phagocyte along with complement factor 6-depleted pooled normal human serum. In addition, we will use flow cytometry platforms employed in our laboratory in order to develop a high-throughput quantitative assay to measure OPA-dependent binding and internalization of N. gonorrhoeae. Overall, results from Core D will help the GV CRC and the field in general to establish the correlate(s) of protection for vaccines against gonorrhea, which to date are poorly understood. When integrated with results from other Cores in the GV CRC, the findings from Core D will contribute to selection of the most promising antigen(s) and platform(s) for the advancement of a novel vaccine for gonorrhea towards licensure.

项目摘要/摘要 核心D：功能抗体研究核心的总体目标是提供一致的， 经过实验控制和验证的平台，用于评估疫苗接种后功能抗体反应 通过淋病疫苗的项目鉴定出淋病抗原 合作研究中心（GV CRC）。血清杀菌活性和致癌性细胞增多已有 与针对致病性奈瑟氏病的保护性免疫反应有关，并且都取决于抗体 结合细菌表面。评估功能抗体反应的这三个参数是 了解疫苗是否有可能引起保护性免疫。因此，核心D至关重要 GV CRC的成功。核心D将为所有四个研究项目提供服务，并将定期进行互动 与该CRC的其他核心相关，可提供独特但互补的专业知识。对于核心D启用 成功完成GV CRC中项目的完成，我们提出了三个具体目的：1）细菌抗体 表面结合：我们将使用成像流式细胞仪来量化血清中免疫的抗体的能力 小鼠结合完整的淋病链球菌的表面。将评估各种应变背景的细菌。 2） 血清杀菌活性（SBA）：我们将测量免疫小鼠血清中抗体的能力或 人类用脑膜炎链球菌血清群B疫苗免疫，以引起SBA对SBA的影响。 菌株，使用抗体的汇集正常人血清作为活性补体的来源。此外 常规菌落数，我们将使用荧光测定法进行并优化高通量定量测定 代谢染料作为SBA的替代度量。 3）打phagococytic活性（OPA）：我们将测量能力 来自免疫小鼠和脑膜杆菌血清群B-免疫的血清中的抗体以增强 使用HL-60人类寄宿细胞作为吞噬细胞，依赖OPA依赖于淋病的N. gonorrhoeae 补体因子6耗尽的合并正常人血清。此外，我们将使用流式细胞仪平台 在我们的实验室中使用，以开发高通量定量测定法以测量OPA依赖性 淋病猪笼草的结合和内在化。总体而言，核心D的结果将有助于GV CRC和领域 一般建立针对淋病的疫苗保护的相关性，迄今为止很差 理解。当与GV CRC中其他核心的结果集成时，核心D的发现将 有助于选择最有前途的抗原和平台，以提高新型疫苗 用于淋病的许可。