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(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寥寥无几。因此，需要新的疫苗技术或佐剂来刺激粘膜部位的保护性免疫。然而，目前的粘膜佐剂有很大的局限性，包括毒性。我们建议开发/建立一种新的重组、无佐剂的黏膜疫苗平台。以人FcγRI(hfcγRI)为靶点的肺炎球菌表面蛋白A(PspA)抗原(Ag)为靶点的单特异性原型融合蛋白(FP)，展示了无佐剂黏膜疫苗平台的可能性。具体地说，这种原型FP在鼻腔给药(I.N.)时增强了免疫原性和对黏膜肺炎链球菌(Sp)攻击的保护作用。给hFcγRI转基因小鼠，在没有佐剂的情况下。我们现在建议创建一种双靶向(多功能)FP：1)通过FcRN介导的FP跨上皮运输，增强疫苗Ag向鼻相关淋巴组织内表达γRI的抗原提呈细胞(APC)的递送。2)通过增加NAT内APC上HFCγRI的交联度，进一步促进抗原内化、树突状细胞(DC)成熟和抗原提呈/T细胞活化。重要的是，由于目前疫苗研究的重点主要是佐剂发现，最大限度地发挥这个无佐剂疫苗平台的效力将对改变人们对佐剂需求的看法至关重要。为了实现这一点，我们将：目的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。这一新型多功能FP随后将在体外测试其能力，以依次增强Ag的跨上皮转运、Ag内化、DC成熟和Ag呈递/T细胞激活。目的：利用SPPSPA抗原黏膜疫苗模型和表达hFcRn/hFcγRI的小鼠，优化FP平台的免疫原性和保护性。具体地说，我们将比较二价和三价hFcRI靶向FP(加上和减去FcRN靶向成分)I.N.然后将选择最具保护性的FP进行进一步开发，并将与获得许可的Sp疫苗进行免疫力和对多种Sp菌株的保护测试。目标3：利用体外培养的AN 人类鼻道的Transwell模型，由上孔(代表鼻腔)中的人类上皮细胞层和下孔(代表NALT)的人类免疫细胞组成。将检验优化的FP将Ag从上井输送到下井并随后增强下井中的人类T和B细胞反应的能力。抗原特异性抗体反应将通过ELISAT/ELISPOT进行检测。T细胞的激活将通过增加CD25的表达、T细胞的增殖和细胞因子的产生来衡量。拟议的研究将对建立这一创新的(双靶向/多功能)疫苗平台作为一种可行的无佐剂粘膜免疫方法至关重要。此外，它的成功将从根本上改变针对多种传染病病原体的疫苗生产/管理模式。