PROCESSING AND SELECTION OF HIV CTL EPITOPES

HIV CTL 表位的处理和选择

• 批准号: 2068302
• 负责人: Cornelia Bergmann
• 金额: $ 11.44万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2068302
• 关键词: MHC class I antigen; Sindbis virus; antigen presentation; antisense nucleic acid; cytotoxic T lymphocyte; endoplasmic reticulum; human immunodeficiency virus 1; laboratory mouse; molecular cloning; protein sequence; protein transport; tissue /cell culture; transfection; vaccinia virus; virus antigen

The goal of this proposal is to determine the influence of subcellular localization and the flanking amino acids on the presentation of an HIV gp 160 epitope to cytotoxic T lymphocytes (CTL). CTL provide a critical component of the protective immune response. Their major role is the elimination of virus infected cells. A prerequisite for the recognition of infected cells is the intracellular degradation and selective presentation of viral antigens in the form of peptides associated with class I molecules at the cell surface. The limited natural selection of viral CTL epitopes is determined by intra- and extracellular mechanisms that are poorly understood. Recombinant vaccinia and sindbis virus expression systems will be used to express peptides containing a defined CTL epitope from HIV. This epitope was chosen because it is recognized by both human and murine CTL. Using molecular techniques these peptides will be localized to the cytosol and the endoplasmic reticulum (ER). Two membrane bound forms of the peptide, one on the cytoplasmic and the other on the luminal side of the ER will be expressed from minigenes. A cytoplasmic "preprocessed" form of the epitope will be used as control for transport into the ER and assembly with the murine Dd class I molecule. The resulting peptides will be tested for their ability to be processed and presented to gp 160 specific CTL. These studies will provide information on the mechanism(s) and subcellular localization of antigen processing. The influence of flanking amino acid sequences on processing will be analyzed by substitution of the wild-type flanking residues to provide insight into the use of recombinant minigenes as potential vaccines. Finally, the subcellular localized antigens will be used to examine the role of the peptide transporter which is believed to be responsible for entry of peptides into the ER. Their role in antigen processing and transport of epitopes will be investigated by testing the localized peptide antigens in cells exhibiting transporter mutations and by downregulating transporter gene expression in normal cells using antisense technology.

该提议的目的是确定亚细胞的影响 在艾滋病毒的介绍中定位和侧翼氨基酸 GP 160表位对细胞毒性T淋巴细胞（CTL）。 CTL提供了关键 保护性免疫反应的组成部分。 他们的主要角色是 消除病毒感染的细胞。 认可的先决条件 感染细胞的细胞内降解和选择性 以与 I类分子在细胞表面。 有限的自然选择 病毒CTL表位由细胞内和细胞外机理确定 理解很差。 重组疫苗和信德氏病毒 表达系统将用于表达包含定义的肽 艾滋病毒的CTL表位。 之所以选择此表位，是因为它被识别 由人类和鼠CTL。 使用分子技术这些肽 将定位于细胞质和内质网（ER）。 二 肽的膜结合形式，一种在细胞质上，另一个在细胞质上 在急诊室的腔侧将以微型烯表示。 一个 表位的细胞质“预处理”形式将用作对照 用于与鼠DD I类运输到急诊室和组装 分子。 由此产生的肽的能力将测试 处理并呈现给GP 160特定CTL。 这些研究会 提供有关机制和亚细胞定位的信息 抗原加工。 侧翼氨基酸序列对 将通过替代野生型侧面来分析处理 残留物可深入了解重组微型烯的使用 潜在疫苗。 最后，亚细胞局部抗原将是 用于检查肽转运蛋白的作用 负责将肽进入急诊室。 它们在抗原中的作用 表位的加工和运输将通过测试来研究 表现出转运蛋白突变的细胞中的局部肽抗原和 通过下调正常细胞中转运蛋白基因的表达 反义技术。