Role of innate immunity in responding to an alphavirus-based adjuvant

先天免疫在响应基于甲病毒的佐剂中的作用

• 批准号: 7914125
• 负责人: BENJAMIN P STEIL
• 金额: $ 4.76万
• 依托单位: GLOBAL VACCINES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7914125
• 关键词: Adjuvant; Adverse effects; Affect; Agonist; Alphavirus; Antigen Targeting; Antigens; Bone Marrow; Bone Marrow Cells; CD8B1 gene; Cells; DNA; Dendritic Cells; Dengue Virus; Detection; Disease; Drug Formulations; Effectiveness; Event; Freund' s Adjuvant; Genome; Goals; HIV; Hepatitis C virus; Immune; Immune response; Immune system; Immunity; Immunologic Receptors; In Vitro; Individual; Infection; Inflammatory; Interferons; Interleukin-6; Investigation; Knockout Mice; Lead; Life Cycle Stages; Ligands; Measures; Mediating; Mucosal Immune Responses; Mus; Natural Immunity; Pathway interactions; Play; Production; Proteins; Public Health; RNA; RNA replication; Replicon; Research; Role; Small Interfering RNA; T-Lymphocyte; Testing; Toll-like receptors; Transfection; Translations; Tuberculosis; Vaccine Adjuvant; Vaccines; Venezuelan Equine Encephalitis Virus; Virus; Work; activating transcription factor; aluminum sulfate; base; cytokine; design; improved; in vitro Assay; in vivo; lymph nodes; migration; mucosal site; novel vaccines; particle; pathogen; ultraviolet damage

DESCRIPTION (provided by applicant): The goal of our proposal is to understand how alphavirus replicon particles (nVRPs) function as adjuvants. nVRPs stimulate a protective systemic and mucosal immune response when co-delivered with a target antigen even when delivered at a non-mucosal site. nVRPs appear to activate the innate immune response, but it is not known how this occurs. Activation of the innate immune response is a crucial event required for protective immunity. By determining how nVRPs activate the innate immune response, our studies may lead to improved rational design and targeting of new adjuvants (such as nVRPs) that can improve both existing and new vaccines to diseases such as tuberculosis, HIV, dengue virus and hepatitis C virus. In our proposal, we will determine which nVRP components activate the innate immune response. We will test the role of translation and replication of nVRP RNA in activating the innate immune response in murine L929 and bone marrow-derived dendritic cells (BMDCs). If we are able to define a particular component(s) of the nVRP that induces the innate immune response, we will test the ability of this component to function as an adjuvant when co-delivered with a target antigen in mice. In our proposal, we will also investigate the innate immune pathways important for responding to nVRP infection. We will investigate the roles of particular innate immune proteins through siRNA knockdown in L929 cells and BMDCs and through using BMDCs from knockout mice. These cells will be infected with nVRPs and measured for activation of the innate immune response through the detection of activated transcription factors and by measuring pro-inflammatory cytokines and type 1 interferons in supernatants from infected cells. We will also use in vitro assays that mimic activation of CD8 T cells by dendritic cells in the lymph node. After BMDCs are infected with nVRPs they stimulate activation of CDS T cells in vitro and we will investigate which innate immune pathways in BMDCs mediate this activity. Finally, we will test the nVRP adjuvant activity in vivo using mice knocked out for particular innate immune proteins. By determing the nVRP components that activate the innate immune response and which innate immune receptors are important for the nVRP adjuvant effect; our studies may lead to adjuvant formulations that contain a cocktail of molecules that specifically activates one or more pathways of the innate immune response. PUBLIC HEALTH RELAVENCE: We will determine which adjuvant components and innate immune proteins are important for stimulation of a protective immune response. Our findings may lead to improved design of adjuvants that can increase the effectiveness of both existing and new vaccines.

描述（由申请人提供）：我们提案的目标是了解甲病毒复制子颗粒（nVRP）如何作为佐剂发挥作用。当 nVRP 与目标抗原共同递送时，即使在非粘膜部位递送，也会刺激保护性全身和粘膜免疫反应。 nVRP 似乎可以激活先天免疫反应，但尚不清楚这是如何发生的。先天免疫反应的激活是保护性免疫所需的关键事件。通过确定 nVRP 如何激活先天免疫反应，我们的研究可能会改进新佐剂（例如 nVRP）的合理设计和靶向，从而改善针对结核病、艾滋病毒、登革热病毒和丙型肝炎病毒等疾病的现有疫苗和新疫苗。在我们的提案中，我们将确定哪些 nVRP 成分激活先天免疫反应。我们将测试 nVRP RNA 的翻译和复制在激活小鼠 L929 和骨髓源性树突状细胞 (BMDC) 的先天免疫反应中的作用。如果我们能够确定 nVRP 中诱导先天免疫反应的特定成分，我们将测试该成分与靶抗原共同递送至小鼠时作为佐剂的能力。在我们的提案中，我们还将研究对于应对 nVRP 感染很重要的先天免疫途径。我们将通过 L929 细胞和 BMDC 中的 siRNA 敲除以及使用来自敲除小鼠的 BMDC 来研究特定先天免疫蛋白的作用。这些细胞将被 nVRP 感染，并通过检测激活的转录因子以及测量感染细胞上清液中的促炎细胞因子和 1 型干扰素来测量先天免疫反应的激活情况。我们还将使用模拟淋巴结中树突状细胞激活 CD8 T 细胞的体外测定。 BMDC 感染 nVRP 后，它们会在体外刺激 CDS T 细胞的激活，我们将研究 BMDC 中哪些先天免疫途径介导这种活性。最后，我们将使用敲除特定先天免疫蛋白的小鼠来测试 nVRP 佐剂的体内活性。通过确定激活先天免疫反应的nVRP成分以及哪些先天免疫受体对于nVRP佐剂作用很重要；我们的研究可能会产生含有分子混合物的佐剂配方，这些分子可以特异性激活先天免疫反应的一个或多个途径。公共卫生相关性：我们将确定哪些佐剂成分和先天免疫蛋白对于刺激保护性免疫反应很重要。我们的研究结果可能会改进佐剂的设计，从而提高现有疫苗和新疫苗的有效性。