T-Cell Receptor Repertoires in Immune Regulation and Response

免疫调节和反应中的 T 细胞受体库

• 批准号: 8157081
• 负责人: Joshua Milner
• 金额: $ 40.22万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/8157081
• 关键词: Affinity; Allergens; Antigens; Collaborations; DNA Sequence; DNA Sequence Rearrangement; Data; Defect; Disease; Evaluation; Goals; Human; Immune; In Vitro; Individual; Infection; Measurement; Memory; Methods; Patients; Peptides; Recording of previous events; Regulation; T-Cell Receptor; United States National Institutes of Health; response; tool

This is the second report for this project. The project consists of the creation of 2 important tools for measuring T-cell receptor diversity. The first, is the use of high-throughput sequencing to measure the diversity of T-cell receptor rearrangements. The strategies and protocol development are ongoing and being done in collaboration with Danny Douek's lab in the NIH VRC. THe sequencing method, and techniques for analysis are nearly completed and patient samples will soon be analyzed using this method. The second tool is a functional measurement of diversity by stimulating patient T-cell samples with peptide libraries. This project is now very mature, as we are able to measure T-cell responses to random peptides in both the naive and memory compartments. We are now validating this method in patients with restricted repertoires. These two tools will be used to better characterize the TCR diversity of two important patient groups. The first is patients whose infectious and immune dysfunctional history is consistant with a T-cell defect, but when lymphocyte subsets are measured, no T-cell deficiency is noted. We hope to define holes in the TCR repertoires of these patients which could explain their infections. The second is in severely atopic patients, whose repertoires may also have similar "holes" but for whom the remaining reactive T-cells therefore develop an abberantly atopic, as opposed to absent response, to a given antigen. These patient cohorts are being assembled now, and as one might expect there are patients in these cohorts with both atopy and infecious predilection. We also hope to apply T-cell receptor repertoire studies to allergen-reactive T-cells, and have already begun to obtain data regarding the TCR sequences of Th1 versus Th2 allergen reactive T-cells. Furthermore, priming of polyclonal, naive human T-cells in vitro with peptide has never been accomplished. We are now able to do so, which will allow for the evaluation of early T-cell signaling and T helper cell differentiation in a fashion never previously done.

这是该项目的第二个报告。该项目包括创建2种测量T细胞受体多样性的重要工具。首先是使用高通量测序来测量T细胞受体重排的多样性。策略和协议制定正在进行中，并与NIH VRC的丹尼·杜克（Danny Douek）的实验室合作完成。测序方法和用于分析的技术几乎完成，并且将很快使用此方法分析患者样本。第二个工具是通过用肽文库刺激患者T细胞样品来对多样性进行功能测量。现在，该项目非常成熟，因为我们能够测量天真和内存室中对随机肽的T细胞反应。现在，我们正在验证库存受限的患者中。这两个工具将用于更好地表征两个重要患者群体的TCR多样性。第一个是患者的传染性和免疫功能障碍史与T细胞缺陷保持一致，但是当测量淋巴细胞亚群时，未发现T细胞缺乏。 我们希望在这些患者的TCR曲目中定义孔，这可以解释他们的感染。第二个是在严重的特有患者中，其曲目可能也具有相似的“孔”，但其余的反应性T细胞出现了极大地产生的，而不是缺乏反应，而不是对给定的抗原。这些患者队列现在正在组装，正如人们可能期望的那样，这些同类群体中都有特应性和偏爱的患者。我们还希望将T细胞受体曲目研究应用于过敏原T细胞，并已经开始获取有关TH1与Th2过敏原反应性T细胞的TCR序列的数据。此外，从未实现多克隆，天真的人类T细胞的启动。我们现在可以这样做，这将允许以以前从未做过的方式评估早期T细胞信号传导和T辅助细胞分化。