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Small molecules to manipulate peptide binding to MHC class I molecules, an optimized method for the generation of MHC tetramers by peptide exchange

操纵肽与 MHC I 类分子结合的小分子，是通过肽交换生成 MHC 四聚体的优化方法

基本信息

批准号：
310813447
负责人：
Professor Dr. Sebastian Springer
金额：
--
依托单位：
Department of Life Sciences and Chemistry
依托单位国家：
德国
项目类别：
Research Grants (Transfer Project)
财政年份：
2016
资助国家：
德国
起止时间：
2015-12-31 至 2019-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/310813447?language=en
关键词：
Small molecules manipulate peptide binding

项目摘要

MHC (Major Histocompatibility Compex) class I molecules play a pivotal role in the antiviral immune response by presenting viral peptides to cytotoxic T cells at the cell surface. Each T cell recognizes a specific class I-peptide complex and subsequently induces apoptosis of the infected cell. These specific T cells proliferate in the course of an antiviral immune response so that the examination of a patient´s T cell pool allows the identification of the most antigenic peptides offering the possibility to predict the progress of the disease or to determine peptides applicable for immunization.So far, relevant T cells are detected with fluorescently labeled class I tetramers: Four class I molecules are folded with the test peptide of interest, biotinylated, and the class I-peptide complexes are tetramerised via fluorescently labeled streptavidin. A class I tetramer efficiently binds to its specific T cell, which can then be detected and quantified. A special type of class I tetramers are the so-called Streptamers: Here, the class I molecules carry a Strep-tag that binds to a mutagenized streptavidin (StrepTactin) with high-affinity and can be released from StrepTactin by the addition of biotin enabling reversible T cell staining.However, T cell staining with Streptamers is a sophisticated and expensive technique impeding immunological research and medical treatment. The aim of this transfer project is the optimization of T cell detection by an improved Streptamer technology based on an innovation we have developed within the scope of a DFG-funded project. In order to detect T cell populations specific for different class I-peptide complexes, each Streptamer must be generated from scratch with the respective test peptides, a procedure that takes weeks every time it has to be repeated. We, in contrast, will pre-produce master Streptamers with folding peptides, which can be exchanged with the help of dipeptides for a test peptide of interest, thereby reducing the production time of a specific Streptamer to a few hours. This innovation will not only accelerate and cheapen T cell detection but will in the future render immune therapy of viral infections more effective.Our cooperation partner, iba GmbH, is a leading supplier of products and services to the life science community based in Göttingen, Germany, and is especially well known for their Strep-tag technology. In the course of this transfer project, we and iba will together develop a prototype kit that will in the future allow each diagnostic and research lab to individually generate Streptamers with peptides of interest in a time- and cost-saving manner. Iba GmbH will later sell these kits or, alternatively, will offer T cell screenings based on that same assay. In the long term, this project will also strengthen the market position of the German company iba against international competitors, especially US american companies.

MHC（主要组织相容性复合体）I类分子通过在细胞表面将病毒肽呈递给细胞毒性T细胞而在抗病毒免疫应答中起关键作用。每个T细胞识别特定的I类肽复合物，随后诱导受感染细胞的凋亡。这些特异性T细胞在抗病毒免疫应答过程中增殖，因此对患者T细胞库的检查可以鉴定出最具抗原性的肽，从而有可能预测疾病的进展或确定适用于免疫的肽。到目前为止，相关的T细胞是用荧光标记的I类四聚体检测的：将四个I类分子与感兴趣的测试肽折叠，生物素化，并通过荧光标记的链霉亲和素将I类-肽复合物四聚化。I类四聚体有效地结合其特异性T细胞，然后可以检测和定量。一种特殊类型的I类四聚体是所谓的Streptamers：在这里，I类分子携带一个Strep标签，该标签以高亲和力结合诱变的链霉亲和素（StrepTactin），并且可以通过添加生物素从StrepTactin释放，从而实现可逆的T细胞染色。然而，使用Streptamers的T细胞染色是一种复杂且昂贵的技术，阻碍了免疫学研究和医学治疗。该转移项目的目的是通过改进的Streptamer技术优化T细胞检测，该技术基于我们在DFG资助项目范围内开发的创新。为了检测对不同的I类肽复合物具有特异性的T细胞群，必须用相应的测试肽从头开始生成每个Streptamer，每次重复该过程都需要数周时间。相反，我们将预先生产具有折叠肽的主Streptamer，其可以在二肽的帮助下交换为感兴趣的测试肽，从而将特定Streptamer的生产时间缩短至几个小时。这一创新不仅将加速T细胞检测并降低检测成本，而且在未来将使病毒感染的免疫治疗更加有效。我们的合作伙伴iba GmbH是一家位于德国哥廷根的生命科学领域的领先产品和服务供应商，其Strep标签技术尤其闻名。在这个转移项目的过程中，我们和iba将共同开发一个原型试剂盒，未来将允许每个诊断和研究实验室以节省时间和成本的方式单独生成具有感兴趣的肽的Streptamer。Iba GmbH稍后将出售这些试剂盒，或者基于相同的检测方法提供T细胞筛查。从长远来看，该项目还将加强德国公司iba在国际竞争对手，特别是美国公司中的市场地位。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

DOI：
10.1126/sciimmunol.aau9039
发表时间：
2019-07-01
期刊：
SCIENCE IMMUNOLOGY
影响因子：
24.8
作者：
Saini, Sunil Kumar;Tamhane, Tripti;Hadrup, Sine Reker
通讯作者：
Hadrup, Sine Reker

Successive crystal structure snapshots suggest the basis for MHC class I peptide loading and editing by tapasin

DOI：
10.1073/pnas.1807656116
发表时间：
2019-03-12
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Hafstrand, Ida;Sayitoglu, Ece Canan;Achour, Adnane
通讯作者：
Achour, Adnane

High-throughput peptide-MHC complex generation and kinetic screenings of TCRs with peptide-receptive HLA-A*02:01 molecules