PMID: 35638972
TCR-T细胞和表达对应MHC的人肿瘤细胞共培养
a Jurkat T-cell line (abbreviated as Jk) engineered with a T-cell receptor (TCR) clonotype (1G4) specific to NYESO-HLA-A2，The fusion of the NYESO peptide with a flexible linker, β2 microglobulin, another flexible linker, and then the heavy chain of HLA-A2 makes up a single-chain trimer that enhances peptide antigen occupancy and presentation to CD8+ T cells.
Anti-CD3 (Invitrogen, 16-0037-81, RRID:AB_468854) and anti-CD28 (Invitrogen, 16-0289-81, RRID:AB_468926) were added into the culture media at a final concentration of 1 μmol/L (when using human PBMCs). 激活T细胞进入分化。
不同的 T 细胞其 TCR 序列很可能不一样。所以从人体分离出的PBMC包含很多不同类型的T细胞，能特异识别响应特定肿瘤细胞的T细胞就会更少。所以体外实验，都是对细胞模型进行编辑，使T细胞能对应识别对应的肿瘤细胞。
TCR 是一种蛋白受体，它可以结合另一种蛋白配体叫做短肽-主要组织相容性复合物（peptide-major histocompatibility complex, pMHC）。其中 MHC 的多样性有限，每个人体内仅表达有限几种 MHC，然而在人类群体中 MHC 存在多态性。TCR 在识别 pMHC 后，所产生的 TCR 下游信号强度。一般信号强度越高，T 细胞杀死癌细胞的能力越强。
细胞表面蛋白的提取，不能用胰酶消化。
For cultured cell lines, cells were detached from the culture dish using the trypsin-free detachment solution Accutase (BioLegend, 423201) to preserve cell-surface proteins.
PD-L1表面蛋白，如果要检测细胞内的蛋白，The assay was performed as described previously ([8](javascript:;)).In brief, cell-surface PD-L1 was labeled with unconjugated anti–PD-L1 (BioLegend, 329703, RRID:AB_940362) for 1 hour on ice and washed twice to remove unbound antibody.
