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

• 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
• 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

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：F。tuarlarensis是革兰氏阴性类别的细胞内粘膜病原体。细胞免疫是 保护这种生物的保护至关重要，而抗体（ABS）延迟感染的进展。定位 对Ag的细胞（APC）上对FC受体（FCR）的抗原（AG）可以增强体液和细胞免疫。 我们假设靶向传染病Ag，例如灭活的F. tularensis（IFT），以粘膜处于FCR 站点将增强防止粘膜挑战的保护。在AIM 1中，我们将研究预先形成的能力 mab-fift复合物和mAb加上IFT混合物，以增强IFT的结合，内在化和表示 APC至Ag特异性T细胞，启动保护性免疫反应的关键事件。我们将：1）检查 MAB：IFT比对mab-Ift结合，内在化和AG表示的影响； 2）确定， 使用鼠标APC，如果MAB加IFT混合物可以代替预先形成的mab-ift； 3）确定是否 上面的FCR靶向策略还可以增强人类APC的IFT结合，内在化和表现。在 AIM 2，我们将确定在小鼠中，mab-ift复合物和/或mAb加上IFT混合物的能力 I.N.，I.D.，I.M.或S.C.增强对I.N.的保护或I.D挑战与现场F. tularensis，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介导的 在AIM 1中观察到的绑定，内在化和表现与由 I.N。施用的FCR靶向免疫原子； 2）确定CD8和CD4 T细胞，B细胞，FCR，AB，， 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 面对FCR的IFT优先定位于淋巴组织，而不是靶向的IFT； 4）确定是否保护 反对I.N.和I.D.可以在外围（I.D.，I.M.或S.C.）免疫后产生挑战； 5） 确定包含CTB（I.N。）和明矾（I.D.，I.M.或S.C.）是否进一步增强了FCR-产生的保护 靶向免疫原。在AIM 3中，我们将验证该疫苗的灵活性和多病原体潜力 平台。具体而言，我们将生成针对肺炎链球菌的FCR靶向亚基疫苗（FC-PSPA） 大量公共卫生问题的细胞外粘膜病原体。 PSPA是S的表面成分。 肺炎在佐剂存在下产生AB依赖性保护。我们将调查 一单价和多价FC-PSPA偶联物包含可变量PSPA的能力，并管理 通过粘膜和外围路线，防止I.N.肺炎链球菌的挑战。的意义 以上研究是实质的：1）新的且更安全的疫苗平台，这既可以产生体液，又产生 需要细胞免疫； 2）在粘膜疫苗的情况下，后者尤其明显； 3）有 迫切需要针对f菌的有效粘膜疫苗，以及更有效的疫苗 S.肺炎； 4）关于FCR在粘膜免疫中的作用以及保护产生的知识 缺乏针对粘膜病原体。拟议的研究将填补上述所有领域的显着空白。