PDF(Pubmed)
Abstract：
Recent studies highlight the integral role of the interferon gamma receptor (IFNγR) pathway in T cell-mediated cytotoxicity against solid but not liquid tumors. IFNγ not only directly facilitates tumor cell death by T cells but also indirectly promotes cytotoxicity via myeloid phagocytosis in the tumor microenvironment. Meanwhile, full human ex vivo immune checkpoint drug screening remains challenging. We hypothesized that an engineered gamma interferon activation site response element luciferase reporter (GAS-Luc2) can be utilized for immune checkpoint drug screening in diverse ex vivo T cell-solid tumor cell co-culture systems. We comprehensively profiled cell surface proteins in ATCC\'s extensive collection of human tumor and immune cell lines, identifying those with endogenously high expression of established and novel immune checkpoint molecules and binding ligands. We then engineered three GAS-Luc2 reporter tumor cell lines expressing immune checkpoints PD-L1, CD155, or B7-H3/CD276. Luciferase expression was suppressed upon relevant immune checkpoint-ligand engagement. In the presence of an immune checkpoint inhibitor, T cells released IFNγ, activating the JAK-STAT pathway in GAS-Luc2 cells, and generating a quantifiable bioluminescent signal for inhibitor evaluation. These reporter lines also detected paracrine IFNγ signaling for immune checkpoint-targeted ADCC drug screening. Further development into an artificial antigen-presenting cell line (aAPC) significantly enhanced T cell signaling for superior performance in these ex vivo immune checkpoint drug screening platforms.
摘要：
最近的研究强调了干扰素γ受体（IFNγR）途径在T细胞介导的针对固体而非液体肿瘤的细胞毒性中的重要作用。IFNγ不仅直接促进T细胞的肿瘤细胞死亡，而且还通过肿瘤微环境中的骨髓吞噬作用间接促进细胞毒性。同时,完整的人体外免疫检查点药物筛选仍然具有挑战性.我们假设工程化的γ干扰素激活位点反应元件荧光素酶报告基因（GAS-Luc2）可用于多种离体T细胞-实体瘤细胞共培养系统中的免疫检查点药物筛选。我们全面分析了ATCC广泛收集的人类肿瘤和免疫细胞系中的细胞表面蛋白，鉴定那些具有已建立和新型免疫检查点分子和结合配体的内源性高表达。然后我们改造了三种表达免疫检查点PD-L1、CD155或B7-H3/CD276的GAS-Luc2报告肿瘤细胞系。在相关的免疫检查点-配体接合时抑制荧光素酶表达。在存在免疫检查点抑制剂的情况下，T细胞释放IFNγ，激活GAS-Luc2细胞中的JAK-STAT通路，并产生用于抑制剂评估的可量化生物发光信号。这些报告细胞系还检测到旁分泌IFNγ信号用于免疫检查点靶向ADCC药物筛选。进一步发展为人工抗原呈递细胞系(aAPC)显著增强T细胞信号传导,以在这些离体免疫检查点药物筛选平台中获得优异的性能。
