Pathways of Antigen Presentation to CD8 T Cells In Vivo

CD 8 + T细胞是针对某些病毒的抗病毒免疫应答的关键组分。初始cd 8 + T细胞 只有在识别细胞表面上的肽-MHC I类复合物后才能分化为效应细胞。 抗原呈递细胞（APC）。所呈递的肽可以通过免疫反应从蛋白质抗原产生。 体内途径的数量。肽可以由病毒感染的APC内表达的抗原产生， 当呈现被称为直接启动时。或者，肽可以从蛋白质产生，所述蛋白质 在呈递前从其他细胞转移到APC，这一过程称为交叉引发。在多大程度 哪种直接引发或交叉引发有助于体内初始CD 8 + T细胞的活化尚不清楚。的 本项目的总体目标是描述用于产生MHC I类肽的机制 在体内初始CD 8 + T细胞的引发期间的复合物。我们的基本假设是蛋白质的改变 表达可以并且确实将体内抗原呈递引导到直接或交叉引发途径中。到 研究这个问题，我们将使用重组病毒表达多种蛋白抗原，并将分析 在体外和体内，抗原呈递给初始和效应CD 8 + T细胞。 靶向病毒抗原在特定组织中表达对抗原呈递途径的影响 在体内使用。我们将研究CD 8 + T细胞引发普遍表达或表达的免疫应答的时间和效率。 组织靶向抗原。我们还将确定负责通过每条路线进行预充的APC，并检查 这种APC的特性是有效启动所必需的。在目标2中，我们将研究微分 抗原呈递作为病毒基因免疫原性降低的机制， 病毒生命周期此外，通过使用天然抗原的例子穿梭到不同的抗原呈递中， 在体内，我们将比较通过直接或交叉致敏的CD 8 + T细胞致敏的效率与通过直接或交叉致敏的CD 8 + T细胞致敏的效率。 体内制备的抗原量。在目标3中，我们将在体外和体内确定蛋白质的特性， 其在交叉致敏期间优先从病毒感染的细胞转移到APC。划定 在体内控制不同抗原呈递途径的使用的机制将为本发明提供基础。 合理设计疫苗和免疫策略，旨在诱导保护性CD 8 + T细胞。 _业绩地点（组织、城市、州） 米尔顿S.好时医疗中心 宾夕法尼亚州立大学 宾夕法尼亚州好时市 关键人员。参见说明。根据需要使用续页提供所需信息 从首席研究员开始。按字母顺序列出所有其他关键人员，姓在前 名称机构 Christopher C.诺伯里宾州州立医学院 宾夕法尼亚州好时市 披露许可声明仅适用于SBIR/STTR。参见说明。[]是[] PHS 398（Rav. 05/01）第_2_页 格式如下。 在项目中的作用 主要研究者 没有 表格第2页 首席研究项目主任（最后，第一，中间）：诺伯里，克里斯托弗C。 必须在每一打印页和每一续页的顶部提供pdnUNRPRAYATOD项目主管的姓名。 研究资助 目录 页码 首页...... 1 说明，

CD8+ T cells are critical components of the anti-viral immune response to some viruses. Naive CD8+ T cells differentiate into effector cells only after recognition of peptide-MHC Class I complexes on the surface of antigen presenting cells (APC). The peptides presented can be generated from protein antigens via a number of pathways in vivo. Peptides can be produced from antigens expressed within a virus-infected APC, when presentation is termed direct-priming. Alternatively, peptides can be produced from proteins that are transferred from other cells to APC prior to presentation, a process known as cross-priming. The extent to which direct-priming or cross-priming contribute to activation of naive CD8+ T cells in vivo is not known. The overall objective of this project is to delineate the mechanisms used to generate MHC Class I-peptide complexes during priming of naive CD8+ T cells in vivo. Our underlying hypothesis is that alteration in protein expression can, and does, direct in vivo antigen presentation into direct or cross-priming pathways. To examine this issue we will use recombinant viruses to express multiple protein antigens and will analyze antigen presentation to naive and effector CD8+ T cells both in vitro and in vivo.ln Aim 1 we will determine the effects of targeting viral antigen for expression in specific tissues on the pathways of antigen presentation used in vivo. We will examine the timing and efficiency of CD8+ T cell priming to ubiquitously expressed or tissue-targeted antigen. We will also identify the APC responsible for priming via each route, and examine the properties of this APC that are necessary for effective priming. In Aim 2 we will examine differential antigen presentation as a mechanism for the reduced immunogenicity of virus genes expressed late in the virus life cycle. In addition, by using a natural example of antigen shuttled into different antigen presentation ;)athways in vivo we will compare the efficiency of CD8+ T cell priming via direct or cross-priming to the mount of antigen made in vivo. In Aim 3 we will determine, in vitro and in vivo, the characteristics of proteins that are preferentially transferred from virus-infected cells to APC during cross-priming. Delineation of the mechanisms governing the use of different antigen presentation pathways in vivo will provide a basis for the rational design of vaccines and immunotherapeutic strategies aimed at induction of protective CD8+ T cells. _ERFORMANCE SITE(S) (organization, city, state) The Milton S. Hershey Medical Center The Pennsylvania State University Hershey, PA KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information Start with Principal Investigator. List all other key personnel in alphabetical order, last name firsL Name Organization Christopher C. Norbury Penn State College of Medicine Hershey, PA Disclosure Permission StatemenL Applicable to SBIR/STTR Only. See instructions. [] Yes [] PHS 398 (Rav. 05/01) Page _2_ in the format shown below. Role on Project Principal Investigator No Form Page 2 Principal InvestigatodProgram Director(Last, first, middle): Norbury, Christopher C. The name of the pdncipal investigatodprogram director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page .................................................................................................................................................. 1 Description,