Novel AAV-based tools for liver immunology

基于 AAV 的新型肝脏免疫学工具

基本信息

批准号：
8702542
负责人：
Ian NICHOLAS Crispe
金额：
$ 18.15万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8702542
关键词：
Affinity Albumins Antigen Presentation Antigen Presentation Pathway Antigen-Presenting Cells Antigens Apoptosis Apoptotic CD4 Positive T Lymphocytes CD8B1 gene Capsid Cause of Death Cell Death Cells Chronic Clonal Expansion Cross Presentation Cross-Priming Development Diphtheria Toxin Disease Effector Cell Engineering Future Goals Hepatitis B Hepatitis C virus Hepatocyte Human Immune response Immune system Immunity Immunology Infection Interferon Type I Interferon-alpha Light Liver Location Malaria Mediating Memory Microscopy Mus Natural Immunity Outcome Ovalbumin Ovum Peptides Population Proteins Recombinant adeno-associated virus (rAAV)Reporter Research Series Serotyping Signal Transduction Site T cell response T memory cell T-Cell Activation T-Lymphocyte Techniques Testing Tropism Vaccines Variant adeno-associated viral vector antigen processing base burden of illness cellular transduction density diphtheria toxin receptor fluorophore gene therapy improved in vivo knock-down liver infection novel pathogen promoter public health relevance small hairpin RNA tool uptake vaccine development vector

项目摘要

DESCRIPTION (provided by applicant): The liver is a site at which antigen encounter can result in either T cell immunity or tolerance, but the influences that determine the outcome of T cell priming are not well understood. In this exploratory/developmental proposal, we will create a set of novel tools based on recombinant adeno- associated virus (rAAV) to explore this issue. Specifically, we will modify the rAAV capsid and promoter to alter cell tropism, we will engineer in a series of fluorescent reporters that will contrast with liver autofluorescence, and we will buld vectors using either a high-affinity antigenic peptides, or sequence variants, or a control peptide Other vectors will promote or inhibit direct antigen presentation by the transduced hepatocytes, or cause the death of transduced hepatocytes thus rendering their antigens available for cross-presentation. Using all of these tools, we will test the impact of antigen density, affinity, cell ype-specific expression and direct versus cross-presentation on primary CD8+ and CD4+ T cell activation, on the development of effector cells, and on memory. These studies will create novel tools of wider usefulness. In particular, the red fluorophores such as mCherry are optimized for in vivo microscopy, Furthermore, if in this study we are able to define optimized priming conditions for memory T cell priming, in future studies we will engineer in pathogen-encoded antigens to test whether AAV could be used as a vaccine vehicle against hepatocellular pathogens.

描述（由申请人提供）：肝脏是抗原相遇可以导致T细胞免疫或耐受性的部位，但是确定T细胞启动结果的影响尚不清楚。在此探索性/发展建议中，我们将创建一组基于重组腺相关病毒（RAAV）的新型工具，以探索这个问题。具体而言，我们将修改Raav Capsid和启动子来改变细胞对tropism，我们将在一系列荧光记者中进行工程，这些记者将与肝自动荧光相反，并且我们将使用高亲和力的抗原肽或其他序列的替代剂或对照肽的抗脉冲或对照抗衡剂的侵蚀，我们将使用高亲和力的抗原性肽或对照抗衡的侵蚀剂的侵蚀或抗衡剂的直接抗衡性或抗衡剂的侵蚀。转导的肝细胞的死亡使其抗原可用于交叉呈递。使用所有这些工具，我们将测试抗原密度，亲和力，细胞特异性表达以及直接与跨呈直接呈现对主要CD8+和CD4+ T细胞激活，对效应细胞发展以及记忆的影响。这些研究将创建新颖的更广泛有用的工具。特别是，针对体内显微镜（此外，如果在这项研究中我们能够为记忆T细胞启动的优化启动条件定义了优化的启动条件，我们将在未来的研究中，我们将在病原体编码的抗原中设计AAV来测试是否可以用作AAV作为AAV的AAV来使用AAV，否则将对AAV进行设计优化的启动条件，以测试是否可以用作A疫苗疫苗，因此我们将在本研究中定义优化的启动条件，从而优化了红色的荧光团（例如MCHERRY）。