Tetramer Core

四聚体核心

• 批准号: 9089837
• 负责人: THOMAS C GREENOUGH
• 金额: $ 27.19万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9089837
• 关键词: Antigens; Avidity; Binding; Biological Assay; CD8B1 gene; Column Chromatography; Complex; Detection; Development; Disease Outcome; Epitopes; Equipment and supply inventories; Evolution; Flow Cytometry; Fluorochrome; Funding; Goals; Human Herpesvirus 4; Immune; Infection; Instruction; Label; MHC Class I Genes; Maintenance; Measurement; Memory; Modeling; Pathology; Peptide/MHC Complex; Peptides; Quality Control; Reagent; Sorting - Cell Movement; Staining method; Stains; T cell response; T-Lymphocyte; TCR Activation; Techniques; Testing; Virus; Virus Diseases; Virus Replication; Western Blotting; cost effective; cross reactivity; improved; programs; response

Multimeric peptide-MHC Class I (pMHCI) reagents enable the detection, characterization and purification of epitope-specific CD8+ T cells by flow cytometry and have greatly facilitated studies to advance our understanding of virus-specific CD8+ T cells. Studies that examine the development and evolution of virus- specific and cross-reactive CD8+ T cells are an important focus of this Program Project. Project I (Luzuriaga) will use the Epstein Barr virus (EBV) model of persistent viral infection to characterize the lineage relationship between effector and memory responses and the factors that influence the evolution of antigen-specific CD8+ T cell responses into the memory CD8+ T cell repertoire. Project 2 (Selin) will determine how cross-reactivity impacts CD8+ T cell selection and function, and influences disease outcome. The Tetramer Core will produce the large quantities of high-quality reagents crucial for both of these projects in a timely and cost effective manner. We utilize well-established techniques for the expression and purification of MHC I molecules, refolding with peptide epitopes, and conjugation with fluorochromes. We plan to expand our inventory to include additional pMHCI reagents that will enable a broader analysis of EBV-specific CD8+ T cell responses and will utilize newer fluorescent labels, such as QDots, to facilitate the simultaneous analyses of multiple EBV epitope-specific CD8+ T cell responses by multiparameter flow cytometry. Over the past funding period, we have developed reagents with altered CD8 binding capacity to better define differences in avidity of T cell receptor interactions with cognate antigen. We will also develop bi-specific heterodimers for characterizing cross-reactive CD8+ T cells.

多聚体肽MHC I类（PMHCI）试剂可以检测，表征和纯化 表位特异性CD8+ T细胞通过流式细胞仪，并具有极大的促进研究以推动我们的 了解病毒特异性CD8+ T细胞。研究病毒的发展和演变的研究 特定和交叉反应的CD8+ T细胞是该程序项目的重要重点。项目i （Luzuriaga）将使用Epstein Barr病毒（EBV）持续病毒感染模型来表征 效应子与记忆反应之间的谱系关系以及影响的因素 抗原特异性CD8+ T细胞响应对存储器CD8+ T细胞库的响应。项目2（Selin）将 确定交叉反应性如何影响CD8+ T细胞的选择和功能，并影响疾病结果。 四聚体核心将产生大量的高质量试剂，这两个项目至关重要 及时有效的方式。我们利用公认的技术来表达和 MHC I分子的纯化，用肽表位重折叠，并与荧光染料结合。我们 计划扩大我们的库存，以包括其他PMHCI试剂，以更广泛的分析 EBV特异性CD8+ T细胞响应，并将利用较新的荧光标签（例如QDOTS）来促进 通过多参数流的多种EBV表位特异性CD8+ T细胞响应的多个EBV表位特异性CD8+ T细胞响应的同时分析 细胞仪。在过去的资金期间，我们开发了具有改变CD8结合能力的试剂 更好地定义了T细胞受体相互作用与同源抗原的差异。我们还将发展 用于表征交叉反应性CD8+ T细胞的双特异性异二聚体。