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

描述（由申请方提供）：本申请中提出的研究将评价天然肽产品在使用鼻内粘膜免疫的无针方法递送时的佐剂活性。我们将建立在以前发表的研究，从我们的小组和利用炭疽重组保护抗原（rPA）和肽免疫原，以评估天然肽血管紧张素-（1-7）（Ang-（1-7））提供鼻佐剂活性以诱导包括血清IgG的血清和粘膜抗原特异性体液和细胞介导的免疫应答的能力，粘膜伊加以及全身和粘膜表位特异性CD 8应答。我们还将启动研究，以更好地确定这种天然肽佐剂的作用机制。我们的具体目标如下所列，并将利用鼻免疫的小鼠模型进行早期临床前研究，以确定与金标准粘膜佐剂霍乱毒素（霍乱毒素由于许多毒性而不适合用于人类）相比，Ang-（1-7）作为鼻疫苗佐剂的安全性和有效性。通过完成本申请中提出的研究所获得的信息将为开发用作无针粘膜疫苗佐剂的新型天然肽提供基础。我们的具体目标是：目标1。评价Ang-（1-7）与蛋白质或肽免疫原经鼻给药时的鼻佐剂活性。为此目的进行的研究将评价当与蛋白质抗原（炭疽保护性抗原）或肽免疫原（C4-V3 HIV-1肽）一起经鼻递送时Ang-（1-7）的佐剂活性。当使用蛋白抗原时，我们将监测疫苗诱导的抗原特异性血清IgG、伊加和IgE应答以及粘膜伊加。当使用肽抗原时，我们将使用IFN 3 ELISPOT和四聚体测定来监测表位特异性CD 8应答的诱导。将Ang-（1-7）的佐剂活性与对照佐剂霍乱毒素和CpG进行比较。目标2.进行研究以评估Ang-（1-7）的作用机制及其用作鼻疫苗佐剂时的安全性。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MAS受体是否是Ang-（1-7）的佐剂活性所必需的，并确定使用Ang-（1-7）是否影响鼻腔接种后鼻腔淋巴组织的细胞组成。其他研究将评估Ang-（1-7）用作鼻疫苗佐剂时的安全性。

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.