CTL RESPONSES TO ADENOVIRUS INFECTIONS IN HUMANS

CTL 对人类腺病毒感染的反应

• 批准号: 2649917
• 负责人: GEOFFREY R KITCHINGMAN
• 金额: $ 17.53万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2649917
• 关键词: Adenoviridae; MHC class I antigen; MHC class II antigen; cellular immunity; cytotoxic T lymphocyte; cytotoxicity; epitope mapping; fibroblasts; gene expression; genetic promoter element; human tissue; molecular cloning; mutant; northern blottings; protein sequence; tissue /cell culture; transfection /expression vector; vaccinia virus

The long-range goal of the work described in this proposal is to obtain an understanding of the cellular immune response in humans to infection by adenovirus. Adenovirus is a frequent cause of infections in children, much less so in adults. Suprisingly little is known about the immune response to adenovirus in humans, however, especially the cellular immune response. We have constructed an in vitro system for the generation of a primary cellular immune response to adenovirus and wish to exploit this system to understand the basic immunology of the response. With this in mind, we propose the following specific aims: 1. Test the hypothesis that multiple epitopes within several different adenovirus proteins are the targets of human CTLs, and that these targets vary depending on HLA type. The amino acids within the Ad5 protein(s) that constitute recognition domains will be determined through the use of viral mutants, expression constructs and peptides. These studies will be performed with CTLs from five individuals of differing HLA type to determine the extent to which the epitopes vary depending on which class I and class II molecules are expressed. 2. Test the hypothesis that the epitopes in adenovirus proteins recognized by CTLs will vary depending on the HLA type of the individual. Viral epitopes recognized by various class I and class II HLA molecules will be determined through the analysis of CTL clones. Peptides representing viral epitopes defined in the first specific aim will be fed to target cells matched at one or more loci, and killing by CTL clones will be determined. 3. Test the hypothesis that different subgroups of adenovirus will have some unique, and some similar, CTL epitopes. The epitopes from adenovirus types 11 (subgroup B) and 31 (subgroup A) that elicit CTL responses will be determined to ascertain whether similar or different regions of the viral proteins are targeted. Successful completion of these aims will provide significant new insight into the cellular immune response to adenovirus infections in humans. The practical application of this knowledge could lead to the development of strategies for the toleraization of individuals to Ad5 to increase the efficacy of gene therapy, and to the development of immunotherapies for immunocompromised individuals with life-threatening adenovirus infections.

这项提案中描述的工作的长期目标是 了解人类的细胞免疫反应 以致腺病毒感染。腺病毒是一种常见的 儿童感染，成人感染更少。令人惊讶的是 对腺病毒的免疫应答知之甚少。 然而，人类，特别是细胞免疫反应。我们 已经构建了一个体外系统来产生一种 对腺病毒的初步细胞免疫应答并希望 利用这个系统来了解猪的基本免疫学 回应。考虑到这一点，我们提出以下具体建议 目标： 1.检验几个表位中有多个表位的假设 不同的腺病毒蛋白是人CTL的靶标，并且 这些靶点因人类白细胞抗原类型不同而不同。氨基酸 在构成识别结构域的Ad5蛋白(S)内 将通过使用病毒突变体、表达 构造物和多肽。这些研究将在以下情况下进行 来自五个不同人类白细胞抗原类型个体的CTL以确定 表位的不同程度取决于I类和 表达II类分子。 2.验证腺病毒蛋白中的表位 CTL识别的基因将根据人类白细胞抗原的类型而有所不同 个人的。各种I类和多类病毒识别的病毒表位 人类白细胞抗原II类分子将通过分析确定 CTL克隆。中定义的代表病毒表位的肽 第一个特定目标将被馈送到与一个目标细胞匹配的目标细胞 或更多的位点，以及CTL克隆的杀伤力将被确定。 3.验证腺病毒不同亚群的假设 将有一些独特的，也有一些相似的CTL表位。这个 腺病毒11型(B亚组)和31型(亚组)的表位 A)将确定引起CTL反应以确定 无论病毒蛋白的相似或不同区域是 有针对性的。 成功完成这些目标将提供重大的新的 对腺病毒的细胞免疫应答的洞察 对人类的感染。这一点的实际应用 知识可以导致制定战略，以 个体对Ad5的耐受性以增加其疗效 基因疗法，以及免疫疗法的发展 携带危及生命的腺病毒的免疫功能受损的人 感染。