Mechanisms of STING-Mediated Mucosal Vaccine Adjuvant Activity of Cyclic di-GMP

STING介导的环二-GMP粘膜疫苗佐剂活性机制

基本信息

批准号：
8880430
负责人：
LEI JIN
金额：
$ 32.99万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-17 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8880430
关键词：
Address Adjuvant Adopted Advanced Development Aluminum Antibody Formation Antigens Area Attenuated Bacteria Binding Biochemical Biological CD4 Positive T Lymphocytes Cell Maturation Cytoplasmic Tail DNA DNA Vaccines DNA Viruses Data Dendritic Cells Dendritic cell activation Development Drug Formulations Equilibrium Europe Exhibits Future Gene Expression Generations Genes Goals Growth Health Host Defense Human IFNAR1 gene IRF3 gene Immune Immune response Immunity Immunization In Vitro Infection Infectious Agent Influenza A Virus, H5N1 Subtype Interferons Interleukin-1 Interleukin-2 Interleukin-6 Knowledge Lead Life MF59 Mediating Molecular Mucosal Immune Responses Mucosal Immunity Mus Production Research Role Route Safety Salts Signal Pathway Signal Transduction Streptococcus Structure Surface TBK1 gene TNF gene Tissues Toxic effect Transducers Up-Regulation Vaccine Adjuvant Vaccines Virulence Wild Type Mouse adaptive immunity base chemokine cytokine immunogenic immunogenicity improved in vivo mucosal vaccination mucosal vaccine novel pathogen response sensor small molecule

项目摘要

DESCRIPTION (provided by applicant): The objective of this proposal is to uncover the in vivo mechanisms by which STING (Stimulator of interferon genes) mediates the mucosal vaccine adjuvant activity of cyclic di-GMP (CDG). Protective mucosal immune responses are most effectively induced by mucosal immunization. However, most of the currently approved human vaccines are administered systemically and generally fail to elicit effective mucosal immunity. Live attenuated mucosal vaccines present safety and acceptability issues while purified antigens are generally poor immunogenics when administered by the mucosal route. CDG exhibits potent mucosal immunogenicity thus, has been explored as a promising mucosal vaccine adjuvant. The mechanism by which CDG executes its mucosal adjuvant activity is unknown, which hinders the further development of CDG as an efficacious mucosal adjuvant. STING, also known as MPYS/MITA, is essential for type I IFN (IFN-I) production by cytosolic sensing of DNA. Recently, we showed that STING mediates IFN-I production by CDG. Subsequent structure studies found that the cytoplasmic tail of STING binds to CDG. It is proposed that STING is a direct sensor for CDG, leading to IFN-I production. However, the in vivo role of STING in CDG-induced vaccine adjuvant activity is not known. In this proposal, we found that STING-/- mice fail to generate antigen-specific antibody response after intranasal immunization of an antigen and CDG. Furthermore, the production of Th1/Th2/Th17 cytokines, proinflammatory cytokines and IFN-I are absent in CDG immunized STING-/- mice. Surprisingly, we found that IFN-I signaling is NOT required for the mucosal adjuvant activity of CDG because IFNAR1-/- mice have the same antigen-specific antibody response as the wild-type mice. This is distinct from STING-mediated DNA vaccine adjuvant activity, which requires IFN-I signaling. We propose two specific Aims to address the in vivo cellular and in vitro molecular mechanisms underlying STING-mediated CDG mucosal adjuvant activity. Aim 1: Dendritic Cells (DC) Expression of STING is essential for the mucosal vaccine adjuvant activity of CDG in vivo. We will use conditional STING-/- and conditional STINGwt/wt mice generated by us to determine the cellular mechanism of STING-mediated CDG adjuvant activity in vivo; Aim 2: Determine the IFN-I stimulation independent mechanism underlying STING-mediated DC activation by CDG. We found that CDG promotes STING-dependent but IFN-I signaling-independent DC maturation. We will use biochemical and molecular biological approaches to identify the proximal signaling transducer of STING that mediates this IFN-I stimulation-independent function. We will further verify its importance in the adjuvant activity of CDG in vivo using KO mice. Currently, there is no vaccine formulation containing a mucosal adjuvant approved for human use. The knowledge generated by our studies can help advance the development of CDG as an efficacious mucosal vaccine adjuvant for human use.

描述（由申请人提供）：该提案的目的是揭示刺痛（干扰素基因刺激剂）介导环状DI-GMP（CDG）的粘膜疫苗辅助活性的体内机制。保护性粘膜免疫反应最有效地由粘膜免疫诱导。但是，大多数当前批准的人类疫苗是系统地施用的，通常无法引起有效的粘膜免疫。通过粘膜途径施用，纯化的抗原的活粘膜疫苗呈现安全性和可接受性问题，而纯化的抗原通常是不良的免疫原性。因此，CDG表现出有效的粘膜免疫原性，已被探讨为有希望的粘膜疫苗辅助。 CDG执行其粘膜辅助活性的机制尚不清楚，这阻碍了CDG作为有效的粘膜佐剂的进一步发展。 sting，也称为MPY/MITA，对于通过DNA的胞质感应而产生的I型IFN（IFN-I）至关重要。最近，我们表明STING介导了CDG的IFN-I产生。随后的结构研究发现，刺的细胞质尾巴与CDG结合。有人提出，Sting是CDG的直接传感器，导致IFN-I产生。然而，尚不清楚Sting在CDG诱导的疫苗辅助活性中的体内作用。在此提案中，我们发现刺痛 - / - 小鼠在抗原和CDG的鼻内免疫后无法产生抗原特异性抗体反应。此外，在CDG免疫刺痛 - / - 小鼠中不存在Th1/Th2/Th17细胞因子，促炎细胞因子和IFN-I的产生。令人惊讶的是，我们发现CDG的粘膜辅助活性不需要IFN-I信号传导，因为IFNAR1 - / - 小鼠具有与野生型小鼠相同的抗原特异性抗体反应。这与STING介导的DNA疫苗辅助活性不同，这需要IFN-I信号传导。我们提出了两个特定的目的，以解决刺激介导的CDG粘膜辅助活性的体内细胞和体外分子机制。 AIM 1：刺的树突状细胞（DC）表达对于CDG体内的粘膜疫苗辅助活性至关重要。我们将使用条件刺痛 - / - 以及我们生成的条件stingwt/wt小鼠来确定体内刺激介导的CDG辅助活性的细胞机制； AIM 2：确定CDG刺激介导的DC激活的IFN-I刺激独立机制。我们发现CDG促进了依赖于STING的，但IFN-I信号与独立的DC成熟。我们将使用生化和分子生物学方法来识别介导该IFN-I刺激非依赖性功能的sting的近端信号传导传感器。我们将进一步验证其在使用KO小鼠体内CDG活性中的重要性。当前，尚无疫苗配方，其中包含批准用于人类使用的粘膜佐剂。我们的研究产生的知识可以帮助推进CDG作为人类使用的有效粘膜疫苗辅助。