Mechanisms Involved in, and Development of, FcR-Enhanced Mucosal Vaccination

FcR 增强粘膜疫苗的参与机制和开发

基本信息

批准号：
8261081
负责人：
Edmund J Gosselin
金额：
$ 38.47万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-01 至 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8261081
关键词：
Adjuvant Antibodies Antigen Presentation Antigen Targeting Antigen-Presenting Cells Antigens Area B-Lymphocytes Binding CD4 Positive T Lymphocytes CD8B1 gene Categories Cells Cellular Immunity Communicable Diseases Complex Development Dose Event Fc Receptor Flow Cytometry Formaldehyde Francisella tularensis Generations Human Humoral Immunities Immune Immune response Immunity Immunization Immunofluorescence Immunologic Immunoglobulin G Infection Inflammation Interferon Type II Knockout Mice Knowledge Label Life Lymphoid Tissue Measures Mediating Monitor Mono-S Monoclonal Antibodies Mucosal Immune Responses Mucosal Immunity Mus Organism Peripheral Play Production Public Health Radiolabeled Role Route Series Streptococcus pneumoniae Subunit Vaccines Surface T-Lymphocyte Testing Vaccines Virulent Work aluminum sulfate antigen binding base biothreat extracellular flexibility improved in vivo interferon-alpha B mucosal site mucosal vaccination mucosal vaccine neonatal Fc receptor pathogen radiotracer receptor function vaccine development

项目摘要

SUMMARY: F. tularensis is a gram-negative Category A intracellular mucosal pathogen. Cellular immunity is critical for protection against this organism, while antibodies (Abs) delay the progression of infection. Targeting antigen (Ag) to Fc receptors (FcR) on Ag presenting cells (APC) can enhance humoral and cellular immunity. We hypothesize targeting infectious disease Ag, such as inactivated F. tularensis (iFt), to FcR at mucosal sites will enhance protection against mucosal challenge. In Aim 1, we will investigate the ability of preformed mAb-iFt complexes and mAb plus iFt mixtures, to enhance binding, internalization, and presentation of iFt by APC to Ag-specific T cells, key events in initiating a protective immune response. We will: 1) Examine the impact of mAb:iFt ratio on mAb-iFt-binding, internalization, and Ag presentation by mouse APC; 2) Determine, using mouse APC, if mAb plus iFt mixtures can be used in place of preformed mAb-iFt; 3) Determine if the above FcR-targeting strategies also enhance IFt-binding, internalization, and presentation by human APC. In Aim 2, we will determine in mice, the ability of mAb-iFt complexes and/or mAb plus iFt mixtures administered i.n., i.d., i.m., or s.c. to enhance protection against i.n. or i.d challenge with live F. tularensis, and identify the humoral and/or cellular components critical to the observed protection. We will: 1) Verify optimal FcR-mediated binding, internalization, and presentation observed in Aim 1, correlates with optimal protection generated by FcR-targeted immunogens administered i.n.; 2) Determine the role of CD8 and CD4 T cells, B cells, FcR, Ab, and IFN-gamma in FcR-dependent protection, using mice lacking these immune components; 3) Determine if FcR-targeted iFt preferentially localizes to lymphoid tissues, versus non-targeted iFt; 4) Determine if protection against i.n. and i.d. challenge can be generated following peripheral (i.d., i.m., or s.c.) immunization; 5) Determine if inclusion of CTB (i.n.) and Alum (i.d., i.m., or s.c.) further enhances protection generated by FcR- targeted immunogens. In Aim 3, we will validate the flexibility and the multi-pathogen potential of this vaccine platform. Specifically, we will generate an FcR-targeted subunit vaccine (Fc-PspA) against S. pneumoniae, an extracellular mucosal pathogen of significant public health concern. PspA is a surface component of S. pneumoniae, which generates Ab-dependent protection in the presence of adjuvant. We will investigate the ability of mono- and multivalent Fc-PspA conjugates containing variable amounts of PspA, and administered via mucosal and peripheral routes, to protect against i.n. challenge with S. pneumoniae. The significance of the above studies is substantial: 1) New and safer vaccine platforms, which generate both humoral and cellular immunity are needed; 2) The latter is particularly evident in the case of mucosal vaccines; 3) There is an urgent need for an effective mucosal vaccine against F. tularensis, and a more efficacious vaccine against S. pneumoniae; 4) Knowledge regarding the role of FcR in mucosal immunity, and the generation of protection against mucosal pathogens, is lacking. The proposed studies will fill significant gaps in all the above areas.

摘要：土拉弗朗西斯菌是一种革兰氏阴性 A 类细胞内粘膜病原体。细胞免疫是对于抵御这种生物体至关重要，而抗体 (Abs) 则可以延缓感染的进展。瞄准抗原（Ag）与Ag呈递细胞（APC）上的Fc受体（FcR）结合可以增强体液和细胞免疫。我们假设将传染病 Ag（例如灭活的土拉弗朗西斯菌 (iFt)）靶向粘膜上的 FcR 位点将增强对粘膜挑战的保护。在目标 1 中，我们将调查执行的能力 mAb-iFt 复合物和 mAb 加 iFt 混合物，通过以下方式增强 iFt 的结合、内化和呈递 APC 到 Ag 特异性 T 细胞，这是启动保护性免疫反应的关键事件。我们将：1）检查 mAb:iFt 比例对 mAb-iFt 结合、内化和小鼠 APC 呈递 Ag 的影响； 2）确定，使用小鼠 APC，如果可以使用 mAb 加 iFt 混合物代替预制的 mAb-iFt； 3) 确定是否上述 FcR 靶向策略还可增强 IFt 结合、内化和人类 APC 的呈递。在目标 2，我们将确定在小鼠中施用 mAb-iFt 复合物和/或 mAb 加 iFt 混合物的能力 i.n.、i.d.、i.m. 或 s.c.加强对 i.n. 的防护或用活土拉弗朗西斯进行 i.d 挑战，并确定对观察到的保护至关重要的体液和/或细胞成分。我们将： 1) 验证最佳 FcR 介导目标 1 中观察到的结合、内化和呈递与由 FcR靶向免疫原注射给药； 2）确定CD8和CD4 T细胞、B细胞、FcR、Ab、和 IFN-γ 在 FcR 依赖性保护中，使用缺乏这些免疫成分的小鼠； 3）确定是否与非靶向 iFt 相比，FcR 靶向 iFt 优先定位于淋巴组织； 4）判断是否保护反对 i.n.和内径外周（腹内、肌内或皮下）免疫后可产生攻击； 5）确定包含 CTB（i.n.）和 Alum（i.d.、i.m. 或 s.c.）是否进一步增强 FcR- 产生的保护靶向免疫原。在目标 3 中，我们将验证该疫苗的灵活性和多病原体潜力平台。具体来说，我们将生产针对肺炎链球菌的 FcR 靶向亚单位疫苗 (Fc-PspA)，引起重大公共卫生问题的细胞外粘膜病原体。 PspA 是 S 的表面成分。肺炎链球菌，在佐剂存在下产生抗体依赖性保护。我们将调查含有不同量 PspA 的单价和多价 Fc-PspA 缀合物的能力，并施用通过粘膜和外周途径，以防止体内感染。肺炎链球菌挑战。意义上述研究是实质性的：1）新的、更安全的疫苗平台，它产生体液和需要细胞免疫； 2) 后者在粘膜疫苗的情况下尤其明显； 3）有迫切需要一种有效的粘膜疫苗来对抗土拉弗氏菌，以及一种更有效的疫苗肺炎链球菌； 4）关于FcR在粘膜免疫中的作用以及保护的产生的知识缺乏针对粘膜病原体的作用。拟议的研究将填补上述所有领域的重大空白。