Functional Antibody Study

功能性抗体研究

• 批准号: 10362592
• 负责人: Alison K Criss
• 金额: $ 21.75万
• 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 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10362592
• 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; Individual; 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; Time; Universities; Vaccination; Vaccine Antigen; 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.

项目总结/摘要 核心D：功能性抗体研究核心的总体目标是提供一致的， 用于评价接种后功能性抗体应答的实验控制和验证的平台 淋病疫苗项目中鉴定的淋病奈瑟菌抗原 合作研究中心（GV CRC）。血清杀菌活性和调理吞噬作用已被证实 参与针对致病性奈瑟氏菌的保护性免疫应答，两者都依赖于抗体 与细菌表面结合。功能性抗体应答的这三个参数的评价是 这对于了解疫苗是否有可能引发保护性免疫至关重要。因此，核心D至关重要 为GV CRC的成功做出贡献。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 与本CRC的其他核心，提供独特但互补的专业知识。对于核心D， 在GV CRC项目的成功完成后，我们提出了三个具体目标：1）细菌抗体 表面结合：我们将使用成像流式细胞术来定量免疫小鼠血清中抗体的能力。 小鼠结合到完整的N.淋病将评价不同菌株背景的细菌。（二） 血清杀菌活性（SBA）：我们将测量来自免疫小鼠的血清中抗体的能力，或 用N.脑膜炎血清群B疫苗，以引发针对一组脑膜炎奈瑟菌的SBA。淋病 菌株，使用抗体耗尽的汇集的正常人血清作为活性补体的来源。除了 传统的菌落计数，我们将试点和优化高通量定量测定使用荧光 代谢染料作为SBA的替代量度。3)调理吞噬活性（OPA）：我们将测量 免疫小鼠血清中的抗体和N.脑膜炎血清群B免疫的人，以增强 OPA依赖的N.淋病，使用HL-60人早幼粒细胞作为吞噬细胞沿着 补体因子6耗尽的混合正常人血清。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来测量OPA依赖性 N.淋病总的来说，核心D的结果将有助于GV CRC和外地 一般来说，建立疫苗对淋病的保护作用的相关性，迄今为止， 明白当与GV CRC中其他核心的结果整合时，核心D的结果将 有助于选择最有希望的抗原和平台，以促进新疫苗的发展 以淋病的名义申请执照