Functional Antibody Study

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个一致的， 用于评估疫苗接种后功能性抗体反应的实验控制和验证平台 通过淋病疫苗项目鉴定的淋球菌抗原 合作研究中心(GV CRC)。血清杀菌活性和吞噬细胞功能 与对致病性奈瑟氏菌的保护性免疫反应有关，两者都依赖于抗体 结合在细菌表面。对功能性抗体反应的这三个参数的评估是 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的成功干杯。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 与这一合作伙伴关系的其他核心，提供不同但互补的专业知识。要使Core D启用 成功完成GV CRC的项目，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后血清中抗体的能力 小鼠与完整的淋病奈瑟菌表面结合。不同菌株背景的细菌将被评估。2) 血清杀菌活性(SBA)：我们将测量免疫小鼠或 用B群脑膜炎奈瑟菌疫苗免疫的人诱导针对淋病奈瑟菌群的SBA 菌株，使用抗体耗尽的正常人血清作为活性补体的来源。除了……之外 传统的菌落计数，我们将试验和优化一种高通量的荧光定量分析 代谢性染料作为SBA的替代指标。3)吞噬细胞活性(OPA)：我们将测量 免疫小鼠和脑膜炎奈瑟氏菌B群免疫人血清中的抗体以增强 以HL-60人早幼粒细胞为吞噬细胞的OPA依赖杀灭淋病奈瑟菌 补体因子6耗尽的正常人血清。此外，我们还将使用流式细胞仪平台 为了建立一种高通量定量测定OPA依赖的方法，我们实验室采用了 淋病奈瑟菌的结合和内化。总体而言，核心D的结果将有助于GV CRC和外地 建立迄今为止效果不佳的淋病疫苗的保护性相关性(S) 明白了。当与GV CRC中其他核心的结果相结合时，核心D的结果将 帮助选择最有希望的抗原(S)和平台(S)，以推动新疫苗的发展 因为淋病而申请执照。

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.