Dissecting the Molecular Immunology of T Cell-Aimed Vacc

剖析 T 细胞靶向疫苗的分子免疫学

• 批准号: 6683837
• 负责人: David Frank Stroncek
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6683837
• 关键词: T lymphocyte; active immunization; cell population study; clinical research; genetic library; genetic polymorphism; high throughput technology; histocompatibility antigens; human subject; immunogenetics; immunologic substance development /preparation; immunoregulation; leukocyte antigen typing; microarray technology; neoplasm /cancer; neoplasm /cancer immunotherapy; polymerase chain reaction; serial analysis of gene expression; time resolved data; vaccine development

Human leukocyte antigen (HLA) genes are the most polymorphic genes in the human genome. Knowledge about HLA polymorphism in relation to possible peptide-based, T-cell-restricted vaccination protocols is important for understanding the physiology of T-cell recognition and to improve strategies of T-cell antigen-specific vaccination. During the last few years the HLA Laboratory has developed and perfected techniques for high-resolution typing of HLA class I and class II molecules using polymerase chain reaction (PCR) techniques and more recently robotic sequencing. With these high-resolution methods it has been possible to achieve several categories of results: o Development of antigen identification program based on co-transfection of HLA and cDNA libraries into permissive antigen presenting cells (in this fashion a new epitope for anti-cancer treatment was identified this year in our lab). o Development of a technology for the preparation of epitope/HLA tetrameric complexes for various HLA alleles and various minimal epitopic sequences that can be used for patient monitoring during vaccination as well as sorting of relevant antigen-specific T-cells. o Preparation of a cDNA library of various HLA alleles to be readily available to our lab and the community at large for the previously mentioned purposes. o Development of high-sensitivity and high-through put technology for the in situ monitoring of T-cell responses during vaccination against cancer by measuring serial gene expression levels by quantitative Real-Time PCR and by cDNA microarray technology. With these techniques we are actively investigating variables involved in the algorithm modulating tumor/host interactions in the context of active vaccination protocols.

人白细胞抗原（HLA）基因是人类基因组中最多的多态基因。关于HLA多态性与可能基于肽的T细胞限制疫苗接种方案有关的知识对于理解T细胞识别的生理学和改善T细胞抗原特异性疫苗的策略非常重要。在过去的几年中，HLA实验室使用聚合酶链反应（PCR）技术以及最近的机器人测序开发并完善了HLA I类和II类分子的高分辨率分型技术。通过这些高分辨率方法，可以实现多种结果：o基于HLA和cDNA文库共转染到允许性抗原呈现细胞中的抗原识别计划的开发（以这种方式，我们在实验室中确定了抗癌治疗的新表位）。 o开发用于制备各种HLA等位基因的表位/HLA四聚体配合物以及可在疫苗接种过程中用于患者监测以及相关抗原特异性T细胞的各种最小表观序列的各种最小表现序列。 o为我们的实验室和整个社区提供各种HLA等位基因的cDNA库的准备。 o通过定量实时PCR和cDNA微阵列技术测量串行基因表达水平，开发高敏性和高通用技术，以对针对癌症的T-Cell反应进行原位监测。通过这些技术，我们正在积极研究在主动疫苗接种方案中调节肿瘤/宿主相互作用的算法中涉及的变量。