An Adjuvant-Independent Dual-Targeted (Multi-Function) Mucosal Vaccine Platform

不依赖佐剂的双靶点（多功能）粘膜疫苗平台

• 批准号: 8911997
• 负责人: Edmund J Gosselin
• 金额: $ 21.88万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-15 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8911997
• 关键词: Activities of Daily Living; Adjuvant; Antibody Response; Antigen Presentation; Antigen Targeting; Antigen-Presenting Cells; Antigens; Attenuated; B-Lymphocytes; Binding; Cell Maturation; Cells; Chimeric Proteins; Clinical; Communicable Diseases; Complex; Data; Dendritic Cells; Development; Dose; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Foundations; Francisella tularensis; Future; Generations; Human; IL2RA gene; Immune; Immune response; Immunity; Immunization; In Vitro; Infection; Inflammatory; Licensing; Life; Lung; Lymphoid Tissue; M cell; Measures; Mediating; Modeling; Mono-S; Mucosal Immune Responses; Mucosal Immunity; Mucous Membrane; Mus; Nose; Perception; Peripheral; Predisposition; Production; Protein Binding; Proteins; Publishing; Recombinants; Risk; Route; Serum Albumin; Staging; Streptococcus pneumoniae; System; T cell response; T-Cell Activation; T-Cell Proliferation; T-Lymphocyte; Technology; Testing; Toxic effect; Transgenic Mice; Vaccination; Vaccine Antigen; Vaccine Research; Vaccines; Variant; Viral; Work; base; biodefense; crosslink; cytokine; enzyme linked immunospot assay; immunogenic; immunogenicity; improved; in vitro Model; in vitro testing; infectious disease model; innovation; monolayer; mouse model; mucosal site; mucosal vaccine; neonatal Fc receptor; novel; novel vaccines; pathogen; pneumococcal surface protein A; prevent; protective efficacy; protein function; prototype; public health relevance; response; success; targeted sequencing; trafficking; vaccine development

 DESCRIPTION (provided by applicant): Most human pathogens enter via mucosal routes. Yet, there are very few licensed mucosal vaccines. Thus, there is a need for new vaccine technologies or adjuvants that stimulate protective immunity at mucosal sites. However, current mucosal adjuvants have significant limitations, including toxicity. We propose to develop/establish a novel recombinant, adjuvant-free, mucosal vaccine platform. The potential for an adjuvant-free mucosal vaccine platform is demonstrated using a mono [human FcγRI (hFcγRI)]-specific prototype fusion protein (FP) consisting of pneumococcal surface protein A (PspA) antigen (Ag) targeted to hFcγRI. Specifically, this prototype FP enhances immunogenicity and protection against a mucosal S. pneumoniae (Sp) challenge when administered intranasally (i.n.) to hFcγRI transgenic mice, in the absence of adjuvant. We now propose creating a dual-targeted (multi-functional) FP which: 1) Enhances delivery of vaccine Ag to hFcγRI-expressing Ag presenting cells (APCs) within the nasal-associated lymphoid tissue (NALT), via FcRn mediated transepithelial transport of FP. 2) Further enhances Ag internalization, dendritic cell (DC) maturation, and Ag presentation/T cell activation, via increased hFcγRI crosslinking on APCs within the NALT. Importantly, with the current focus of vaccine research primarily on adjuvant discovery, maximizing the potency of this adjuvant-free vaccine platform will be crucial to changing perceptions regarding the need for adjuvant. To achieve this, we will: Aim 1) Add an FcRn targeting sequence to the prototype (mono-targeted) PspA containing FP, as well as convert the bivalent FP to a trivalent FP. This novel multi-functional FP will then be tested in vitro for its ability to sequentially enhance transepithelial transport of Ag, Ag internalization, DC maturation, and Ag presentation/T cell activation. Aim 2: Optimize FP platform immunogenicity and protective efficacy utilizing the Sp PspA Ag mucosal vaccine model and hFcRn/hFcγRI-expressing mice. Specifically, we will compare bivalent vs. trivalent hFcRI-targeted FP (plus and minus the FcRn targeting component) i.n. The most protective FP will then be selected for further development and will also be tested for immunity and protection against multiple Sp strains versus a licensed Sp vaccine. Aim 3: Utilize an in vitro transwell model of the human nasal tract consisting of a human epithelial cell layer in the top well (representing the nasal lumen) and human immune cells in the lower well (representing the NALT). The ability of the optimized FP to transport Ag from the upper well to the lower well and subsequently enhance human T and B cell responses in the lower well will be examined. Ag-specific antibody responses will be measured by ELISA/ELISPOT. T cell activation will be measured via increased CD25 expression, T cell proliferation, and cytokine production. The proposed studies will be crucial to establishing this novel and innovative (dual-targeted/multi-functional) vaccine platform as a viable adjuvant-free approach for mucosal immunization. Furthermore, its success will fundamentally transform the paradigm for vaccine generation/administration against a wide array of infectious disease agents.

 描述（由申请人提供）：大多数人类病原体通过粘膜途径进入，然而，获得许可的粘膜疫苗非常少，因此需要刺激粘膜部位的保护性免疫的新疫苗技术或佐剂。我们建议开发/建立一种新型重组、无佐剂、粘膜疫苗平台。粘膜疫苗平台使用由针对 hFcγRI 的肺炎球菌表面蛋白 A (PspA) 抗原 (Ag) 组成的单 [人 FcγRI (hFcγRI)] 特异性原型融合蛋白 (FP) 进行验证。具体而言，该原型 FP 增强了免疫原性和针对 hFcγRI 的保护。鼻内（i.n）给予 hFcγRI 转基因小鼠时，在没有我们现在建议创建一种双靶向（多功能）FP，它： 1）通过 FcRn 介导的跨上皮，增强疫苗 Ag 向鼻相关淋巴组织 (NALT) 内表达 hFcγRI 的 Ag 呈递细胞 (APC) 的递送。 2) 通过增加 hFcγRI 交联，进一步增强 Ag 内化、树突状细胞 (DC) 成熟和 Ag 呈递/T 细胞激活。重要的是，目前疫苗研究的重点主要是佐剂的发现，最大限度地发挥这种无佐剂疫苗平台的效力对于改变对佐剂需求的看法至关重要。为了实现这一目标，我们将： 1) 将 FcRn 靶向序列添加到含有 FP 的原型（单靶向）PspA 中，并将二价 FP 转化为三价 FP，然后对这种新型多功能 FP 进行测试。目标 2：利用 Sp PspA Ag 粘膜疫苗模型和 hFcRn/hFcγRI- 优化 FP 平台的免疫原性和保护功效。具体来说，我们将比较二价和三价 hFcRI 靶向 FP（加上和减去 FcRn 靶向成分）。然后，将选择最具保护性的 FP 进行进一步开发，并将针对多种 Sp 菌株与许可的 Sp 疫苗进行免疫和保护测试。 人鼻道的 Transwell 模型由上孔（代表鼻腔）中的人类上皮细胞层和下孔中的人类免疫细胞（代表 NALT）组成。优化后的 FP 从上孔转运 Ag 的能力。将通过 ELISA/ELISPOT 测量 T 细胞活化，并通过增加的 CD25 表达、T 细胞来检查下孔中的人 T 和 B 细胞反应。拟议的研究对于建立这种新颖且创新的（双靶点/多功能）疫苗平台作为可行的无佐剂粘膜免疫方法至关重要。此外，其成功将从根本上改变粘膜免疫的范式。针对多种传染病病原体的疫苗生产/管理。