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：
8289839
负责人：
Ian NICHOLAS Crispe
金额：
$ 23.63万
依托单位：
SEATTLE BIOMEDICAL RESEARCH INSTITUTE
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 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8289839
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 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. PUBLIC HEALTH RELEVANCE: Infections of liver cells impose a large disease burden both in the USA and globally. The way the immune system responds to such infections is poorly understood. In the project we will use techniques from the field of gene therapy to create new tools that will increase understanding of immune responses to infections of the liver.

描述（由申请人提供）：肝脏是抗原相遇可导致 T 细胞免疫或耐受的部位，但决定 T 细胞启动结果的影响尚不清楚。在这个探索性/开发性提案中，我们将创建一套基于重组腺相关病毒（rAAV）的新工具来探索这个问题。具体来说，我们将修改 rAAV 衣壳和启动子以改变细胞向性，我们将设计一系列与肝脏自发荧光形成对比的荧光报告基因，并且我们将使用高亲和力抗原肽或序列变体或对照肽其他载体将促进或抑制转导的肝细胞的直接抗原呈递，或导致转导的肝细胞死亡，从而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、细胞类型特异性表达以及直接与交叉呈递对初级 CD8+ 和 CD4+ T 细胞激活、效应细胞发育和记忆的影响。这些研究将创造出具有更广泛用途的新工具。特别是，mCherry 等红色荧光团针对体内显微镜进行了优化。此外，如果在本研究中我们能够定义记忆 T 细胞启动的优化启动条件，那么在未来的研究中，我们将设计病原体编码抗原来测试是否AAV 可用作对抗肝细胞病原体的疫苗载体。公共卫生相关性：肝细胞感染给美国和全球带来了巨大的疾病负担。人们对免疫系统对此类感染的反应方式知之甚少。在该项目中，我们将使用基因治疗领域的技术来创建新工具，以增进对肝脏感染免疫反应的了解。