胰腺导管腺癌（PDAC）仍然是一种特别具有侵袭性的疾病，有效的治疗方法很少。已知 PDAC 肿瘤免疫微环境 (TIME) 具有免疫抑制作用。溶瘤病毒可以通过免疫原性细胞死亡（ICD）增加肿瘤免疫原性。我们专注于肿瘤选择性 (vvDD) 和细胞因子武装的西储牛痘病毒（vvDD-IL2 和 vvDD-IL15）和感染的癌细胞系以及患者来源的原代 PDAC 细胞。在共培养实验中，我们研究了细胞毒性反应和人类自然杀伤（NK）的激活。通过测量病毒编码的 YFP 来评估感染和病毒复制。然后，我们通过深入的蛋白质组分析、免疫印迹和 TUNEL 测定来分析细胞内信号传导过程和溶瘤作用。将模拟或病毒感染的癌细胞系与同种异体 PBMC 或 NK 细胞系共培养后，分析 CD56 NK 细胞的活化、细胞毒性和效应功能。测量了感染后危险信号的剂量和时间依赖性释放。病毒有效地进入 PDAC 细胞，发出 YFP 信号并导致伴随的溶瘤作用。蛋白质组显示 PDAC 中正常活跃的核心信号通路（例如 MAPK-ERK 信号通路）发生了重编程。感染后释放与危险相关的分子模式，并刺激共培养的 NK 细胞以增强效应细胞毒性。 NK 细胞亚型分析揭示了罕见 CD56dimCD16dim 群体的数量增加和激活。肿瘤细胞杀伤主要是通过 Fas 配体而不是颗粒释放触发的，从而导致显着的细胞凋亡。总体而言，携带细胞因子的痘苗病毒在体外诱导 NK 细胞活化并增强对人 PDAC 细胞的细胞毒性。我们可以证明携带细胞因子的病毒以癌细胞为目标，因此具有调节 PDAC 中 TIME 的巨大潜力。© 2024 作者。约翰·威利出版的《国际癌症杂志》

Pancreatic ductal adenocarcinoma (PDAC) remains a particularly aggressive disease with few effective treatments. The PDAC tumor immune microenvironment (TIME) is known to be immune suppressive. Oncolytic viruses can increase tumor immunogenicity via immunogenic cell death (ICD). We focused on tumor-selective (vvDD) and cytokine-armed Western-reserve vaccinia viruses (vvDD-IL2 and vvDD-IL15) and infected carcinoma cell lines as well as patient-derived primary PDAC cells. In co-culture experiments, we investigated the cytotoxic response and the activation of human natural killer (NK). Infection and virus replication were assessed by measuring virus encoded YFP. We then analyzed intracellular signaling processes and oncolysis via in-depth proteomic analysis, immunoblotting and TUNEL assay. Following the co-culture of mock or virus infected carcinoma cell lines with allogenic PBMCs or NK cell lines, CD56+ NK cells were analyzed with respect to their activation, cytotoxicity and effector function. Both, dose- and time-dependent release of danger signals following infection were measured. Viruses effectively entered PDAC cells, emitted YFP signals and resulted in concomitant oncolysis. The proteome showed reprogramming of normally active core signaling pathways in PDAC (e.g., MAPK-ERK signaling). Danger-associated molecular patterns were released upon infection and stimulated co-cultured NK cells for enhanced effector cytotoxicity. NK cell subtyping revealed enhanced numbers and activation of a rare CD56dimCD16dim population. Tumor cell killing was primarily triggered via Fas ligands rather than granule release, resulting in marked apoptosis. Overall, the cytokine-armed vaccinia viruses induced NK cell activation and enhanced cytotoxicity toward human PDAC cells in vitro. We could show that cytokine-armed virus targets the carcinoma cells and thus has great potential to modulate the TIME in PDAC.© 2024 The Author(s). International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.