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: 7660124
• 负责人: Edmund J Gosselin
• 金额: $ 37.83万
• 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 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7660124
• 关键词: Adjuvant; Antibodies; Antigen Targeting; 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; Monoclonal Antibodies; Mucosal Immune Responses; Mucosal Immunity; Mus; Organism; Peripheral; Play; Production; Public Health; Radiolabeled; Research; Role; Route; Series; Streptococcus pneumoniae; Subunit Vaccines; Surface; T-Lymphocyte; Testing; Vaccines; Virulent; Work; aluminum sulfate; base; biothreat; extracellular; flexibility; improved; in vivo; interferon-alpha B; mucosal site; mucosal vaccination; mucosal vaccine; neonatal Fc receptor; pathogen; public health relevance; radiotracer; receptor function; vaccine development

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The significance of the proposed studies to public health is multi-fold. There is a need for new and safer vaccine platforms, in particular as it applies to mucosal immunity. In addition, an effective mucosal vaccine against F. tularensis (a Category A biothreat agent), and a more efficacious vaccine against S. pneumoniae (a pathogen of significant public health concern), are also needed. We provide strong evidence the proposed studies will satisfy these needs, and fill significant gaps in our knowledge regarding the role and use of Fc receptors in mucosal and peripheral immunity against intracellular and extracellular pathogens.

描述（由申请人提供）：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。细胞免疫对于抵御这种生物体至关重要，而抗体 (Ab) 则可以延缓感染的进展。将抗原 (Ag) 靶向 Ag 呈递细胞 (APC) 上的 Fc 受体 (FcR) 可以增强体液和细胞免疫。我们假设，针对粘膜部位的 FcR 来靶向传染病 Ag，例如灭活的土拉弗朗西斯菌 (iFt)，将增强针对粘膜攻击的保护作用。在目标 1 中，我们将研究预制 mAb-iFt 复合物和 mAb 加 iFt 混合物的能力，以增强 APC 将 iFt 结合、内化和呈递给 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) 使用小鼠 APC 确定是否可以使用 mAb 加 iFt 混合物代替预制的 mAb-iFt； 3) 确定上述 FcR 靶向策略是否也增强人类 APC 的 IFt 结合、内化和呈递。在目标 2 中，我们将在小鼠中确定 mAb-iFt 复合物和/或 mAb 加 iFt 混合物经内、皮内、肌内或皮下施用的能力。加强对 i.n. 的防护或用活土拉弗朗西斯进行 i.d 攻击，并鉴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) 验证目标 1 中观察到的最佳 FcR 介导的结合、内化和呈递，与注射 FcR 靶向免疫原产生的最佳保护相关； 2) 使用缺乏这些免疫成分的小鼠，确定 CD8 和 CD4 T 细胞、B 细胞、FcR、Ab 和 IFN-gamma 在 FcR 依赖性保护中的作用； 3) 确定 FcR 靶向 iFt 是否优先定位于淋巴组织，而不是非靶向 iFt； 4) 确定是否能防止 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 靶向免疫原产生的保护。在目标 3 中，我们将验证该疫苗平台的灵活性和多病原体潜力。具体来说，我们将生产一种针对肺炎链球菌的 FcR 靶向亚单位疫苗 (Fc-PspA)，肺炎链球菌是一种引起重大公共卫生问题的细胞外粘膜病原体。 PspA 是肺炎链球菌的表面成分，在佐剂存在下产生抗体依赖性保护。我们将研究含有不同量 PspA 的单价和多价 Fc-PspA 缀合物，并通过粘膜和外周途径给药，预防静脉内感染的能力。肺炎链球菌挑战。上述研究的意义重大：1）需要新的、更安全的疫苗平台，能够产生体液免疫和细胞免疫； 2) 后者在粘膜疫苗的情况下尤其明显； 3) 迫切需要针对土拉热杆菌的有效粘膜疫苗，以及更有效的针对肺炎链球菌的疫苗； 4）缺乏关于FcR在粘膜免疫中的作用以及针对粘膜病原体的保护作用的知识。拟议的研究将填补上述所有领域的重大空白。公共卫生相关性：拟议研究对公共卫生的意义是多方面的。需要新的、更安全的疫苗平台，特别是因为它适用于粘膜免疫。此外，还需要针对土拉杆菌（A 类生物威胁因子）的有效粘膜疫苗和针对肺炎链球菌（引起重大公共卫生问题的病原体）的更有效的疫苗。我们提供了强有力的证据，所提出的研究将满足这些需求，并填补我们关于 Fc 受体在针对细胞内和细胞外病原体的粘膜和外周免疫中的作用和用途的知识中的重大空白。