A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells
A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells
复制标题
用于大规模检测抗原特异性 T 细胞的灵活 MHC I 类多聚体加载系统
DOI：
10.1084/jem.20180156
复制
发表时间：
2018-05-01
影响因子：
15.3
通讯作者：
Ovaa, Huib
中科院分区：
文献类型：
--
作者：
Luimstra, Jolien J.;Garstka, Malgorzata A.;Ovaa, Huib
适应性免疫是由T细胞识别主要组织相容性复合体（MHC）呈递的特异性抗原而启动的。MHC多聚体技术已被开发用于检测、分离和表征感染、自身免疫和癌症中的T细胞。在这里，我们提出了一个简单，快速，灵活，高效的方法来产生许多不同的MHC I类（MHC I）多聚体在平行使用温度介导的肽交换。我们为HLA-A*02：01和H-2Kb设计了条件肽，它们在低温下形成稳定的肽-MHC I复合物，但在暴露于确定的高温时解离。所得到的条件性MHC I复合物，无论是单独的还是制备为即用型多聚体，都可以在短时间内快速加载所选肽而无需额外处理。我们证明了这种方法的简便性和灵活性，通过监测异基因干细胞移植受体的抗病毒免疫结构，并通过分析CD 8 + T细胞对淋巴细胞性脉络丛脑膜炎病毒或巨细胞病毒感染小鼠的病毒表位的反应。
Adaptive immunity is initiated by T cell recognition of specific antigens presented by major histocompatibility complexes (MHCs). MHC multimer technology has been developed for the detection, isolation, and characterization of T cells in infection, autoimmunity, and cancer. Here, we present a simple, fast, flexible, and efficient method to generate many different MHC class I (MHC I) multimers in parallel using temperature-mediated peptide exchange. We designed conditional peptides for HLA-A*02:01 and H-2Kb that form stable peptide–MHC I complexes at low temperatures, but dissociate when exposed to a defined elevated temperature. The resulting conditional MHC I complexes, either alone or prepared as ready-to-use multimers, can swiftly be loaded with peptides of choice without additional handling and within a short time frame. We demonstrate the ease and flexibility of this approach by monitoring the antiviral immune constitution in an allogeneic stem cell transplant recipient and by analyzing CD8+ T cell responses to viral epitopes in mice infected with lymphocytic choriomeningitis virus or cytomegalovirus.