Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, 72076 Tübingen, Germany.
Cluster of Excellence iFIT (EXC2180) "Image-Guided and Functionally Instructed Tumor Therapies", University of Tübingen, 72076 Tübingen, Germany.
Cells. 2022 Oct 31;11(21):3451. doi: 10.3390/cells11213451.
Elongated peptides (EPs), containing possibly one or multiple epitope/s, are increasingly used for the screening of antigen-specific CD8 and CD4 cell responses. Here, we present an in vitro protocol that allows the amplification of antigen-specific cells and the subsequent functional analysis of both T cell types using EPs. Known viral-derived epitopes were elongated to 20 mer EPs on the N-, C-, and both termini for HLA class I binders, or on the N- and C- termini for HLA class II binders. With EP stimulation only, the percentage of responding CD8 T cells was dependent on the elongation site of the EP, whereas CD4 T cell responses were completely lost in 22% of the tests performed ex vivo. A short-term amplification step plus the addition of a TLR3 agonist (Poly-ICLC) together with an increased EP concentration improved markedly the detection of CD8 and CD4 T cell reactivities.
延伸肽（EPs），包含一个或多个表位/抗原，越来越多地用于筛选抗原特异性 CD8 和 CD4 细胞反应。在这里，我们提出了一种体外方案，允许使用 EPs 扩增抗原特异性细胞，并随后对两种 T 细胞类型进行功能分析。已知的病毒衍生表位被延伸成 20 个氨基酸的 EPs，用于 HLA Ⅰ类结合物的 N-、C-和两端，或用于 HLA Ⅱ类结合物的 N-和 C-末端。仅用 EP 刺激，反应性 CD8 T 细胞的百分比取决于 EP 的延伸部位，而在 22%的体外试验中，CD4 T 细胞反应完全丧失。一个短期的扩增步骤加上添加 TLR3 激动剂（Poly-ICLC）和增加 EP 浓度显著提高了 CD8 和 CD4 T 细胞反应性的检测。
