Evaluation of the nasal adjuvant activity of angiotensin peptide Ang-(1-7)

描述(由申请人提供)：本申请中提出的研究将评估一种天然多肽产品在使用鼻腔粘膜免疫的无针方法给药时的佐剂活性。我们将在我们团队以前发表的研究的基础上，利用炭疽重组保护性抗原(RPA)和多肽免疫原来评估天然多肽Ang-(1-7)(Ang-(1-7))是否能够提供鼻佐剂活性，以诱导血清和粘膜抗原特异性体液和细胞免疫反应，包括血清IgG、黏膜IgA以及全身和粘膜表位特异性CD8反应。我们还将启动研究，以更好地确定这种天然多肽佐剂的作用机制。我们的具体目标如下，将利用小鼠鼻腔免疫模型进行早期临床前研究，以确定Ang-(1-7)作为鼻用疫苗佐剂的安全性和有效性，并与黄金标准的霍乱黏膜佐剂毒素(霍乱毒素由于多种毒性而不适合在人类身上使用)进行比较。本申请中建议的研究的完成所获得的信息将为开发一种用作无针粘膜疫苗佐剂的新型天然多肽提供基础。我们的具体目标是：目的1.评价Ang-(1-7)鼻腔注射蛋白质或多肽免疫原时的佐剂活性。为此目的开展的研究将评估Ang-(1-7)与蛋白质抗原(炭疽保护性抗原)或多肽免疫原(C4-V3 HIV-1肽)鼻腔注射时的佐剂活性。在使用该蛋白抗原时，我们将监测疫苗诱导的抗原特异性血清免疫球蛋白G、免疫球蛋白A和免疫球蛋白E反应以及粘膜免疫球蛋白A。当使用多肽抗原时，我们将使用IFN3 ELISPOT和四聚体分析来监测表位特异性CD8反应的诱导。将Ang-(1-7)的佐剂活性与对照佐剂霍乱毒素和CpG进行比较。目的2.评价血管紧张素-(1-7)作为鼻腔疫苗佐剂的作用机制及其安全性。已知Ang-(1-7)通过MAS受体发出信号。为此目的进行的研究将利用MAS拮抗剂A-779来确定Ang-(1-7)的佐剂活性是否需要MAS受体，并确定Ang-(1-7)的使用是否影响鼻腔免疫后鼻腔淋巴组织的细胞组成。其他研究将评估Ang-(1-7)作为鼻用疫苗佐剂的安全性。 公共卫生相关性：本申请中提出的研究将评估血管紧张素肽Ang-(1-7)作为疫苗佐剂的能力，并增强通过鼻腔途径输送的疫苗的免疫原性。这些疫苗被设计为经鼻腔注射，有朝一日可能会取代针头注射的疫苗。为了使鼻用疫苗的发展取得进展，需要鉴定和开发安全有效的鼻用疫苗佐剂。

DESCRIPTION (provided by applicant): Studies proposed in this application will evaluate the adjuvant activity of a natural peptide product when delivered using the needle-free method of intranasal mucosal immunization. We will build upon previous published studies from our group and utilize anthrax recombinant protective antigen (rPA) and a peptide immunogen to evaluate the natural peptide angiotensin-(1-7) (Ang-(1-7)) for its ability to provide nasal adjuvant activity for the induction of serum and mucosal antigen-specific humoral and cell- mediate immune responses to include serum IgG, mucosal IgA as well as systemic and mucosal epitope-specific CD8 responses. We will also initiate studies to better define the mechanism of action of this natural peptide adjuvant. Our specific aims are listed below and will utilize a mouse model of nasal immunization to perform early-stage preclinical studies to determine the safety and efficacy of Ang-(1-7) as a nasal vaccine adjuvant when compared to the gold-standard mucosal adjuvant cholera toxin (cholera toxin is not suitable for use in humans due to numerous toxicities). Information learned by the completion of the studies proposed in this application will provide a foundation for the development of a novel, natural peptide for use as an adjuvant for needle-free mucosal vaccines. Our specific aims are: Aim 1. Evaluate the nasal adjuvant activity of Ang-(1-7) when nasally delivered with protein or peptide immunogens. Studies performed in this aim will evaluate the adjuvant activity of Ang-(1-7) when nasally delivered with a protein antigen (anthrax protective antigen) or a peptide immunogen (C4-V3 HIV-1 peptide). When using the protein antigen, we will monitor vaccine-induced antigen-specific serum IgG, IgA and IgE responses as well as mucosal IgA. When using the peptide antigen, we will monitor the induction of epitope-specific CD8 responses using IFN3 ELISPOT and tetramer assays. The adjuvant activity of Ang-(1-7) will be compared to control adjuvants cholera toxin and CpG. Aim 2. Perform studies to evaluate the mechanism of action of Ang-(1-7) and its safety when used as a nasal vaccine adjuvant. Ang-(1-7) is known to signal via the MAS receptor. Studies performed in this aim will utilize the MAS antagonist A-779 to determine if the MAS receptor is required for the adjuvant activity of Ang-(1-7) and to determine if the use of Ang-(1-7) influences the cellular composition of the nasal cavity lymphoid tissue after nasal vaccination. Additional studies will evaluate the safety of Ang-(1-7) when used as a nasal vaccine adjuvant. PUBLIC HEALTH RELEVANCE: Studies proposed in this application will evaluate the angiotensin peptide Ang-(1-7) for its ability to be used as a vaccine adjuvant and enhance the immunogenicity of vaccines delivered by the nasal route. These vaccines are designed to be delivered intranasally and may someday replace vaccines injected with a needle. The identification and development of safe and effective nasal vaccine adjuvants is needed to make progress in the development of nasally-administered vaccines.