Department of Gastroenterology, Endocrinology, Infectious Diseases and Metabolism, University Hospital Marburg, Philipps University Marburg, 35043 Marburg, Germany.
Department of Internal Medicine I, University Hospital Ulm, 89081 Ulm, Germany.
Cells. 2023 Jan 31;12(3):456. doi: 10.3390/cells12030456.
In pancreatic ductal adenocarcinoma (PDAC), the infiltration of CD8 cytotoxic T cells (CTLs) is an important factor in determining prognosis. The migration pattern and interaction behavior of intratumoral CTLs are pivotal to tumor rejection. NLRP3-dependent proinflammatory cytokines IL-1β and IL-18 play a prominent role for CTL induction and differentiation. Here, we investigate the effects of T-cellular IL-1R and IL-18R signaling for intratumoral T-cell motility. Murine adenocarcinoma cell line Panc02 was stably transfected with ovalbumin (OVA) and fluorophore H2B-Cerulean to generate PancOVA H2B-Cerulean tumor cells. Dorsal skinfold chambers (DSFC) were installed on wild-type mice, and PancOVA H2B-Cerulean tumor cells were implanted into the chambers. PancOVA spheroids were formed using the Corning Matrigel-based 3D cell culture technique. CTLs were generated from OT-1 mice, OT-1 mice, or OT-1 mice and were marked with fluorophores. This was followed by the adoptive transfer of CTLs into tumor-bearing mice or the application into tumor spheroids. After visualization with multiphoton microscopy (MPM), Imaris software was used to perform T-cell tracking. Imaris analysis indicates a significantly higher accumulation of CTLs in PancOVA tumors and a significant reduction in tumor volume compared to wild-type CTLs. CTLs covered a longer distance (track displacement length) in comparison to wild-type (WT) CTLs, and had a higher average speed (mean track speed). The analysis of instantaneous velocity suggests a higher percentage of arrested tracks (arrests: <4 μm/min) for CTLs. Our data indicate the contribution of IL-18R signaling to T-cell effector strength, warranting further investigation on phenomena such as intratumoral T-cell exhaustion.
在胰腺导管腺癌（PDAC）中，CD8 细胞毒性 T 细胞（CTL）的浸润是决定预后的重要因素。肿瘤内 CTL 的迁移模式和相互作用行为对肿瘤排斥至关重要。NLRP3 依赖性前炎性细胞因子 IL-1β 和 IL-18 对 CTL 的诱导和分化起着突出的作用。在这里，我们研究了 T 细胞 IL-1R 和 IL-18R 信号对肿瘤内 T 细胞迁移的影响。用卵清蛋白（OVA）和荧光蛋白 H2B-Cerulean 稳定转染小鼠腺癌细胞系 Panc02，生成 PancOVA H2B-Cerulean 肿瘤细胞。在野生型小鼠背部皮褶室（DSFC）中安装皮褶室，并将 PancOVA H2B-Cerulean 肿瘤细胞植入皮褶室。使用康宁基于基质胶的 3D 细胞培养技术形成 PancOVA 球体。从 OT-1 小鼠、 OT-1 小鼠或 OT-1 小鼠生成 CTL，并标记荧光蛋白。然后将 CTL 过继转移到荷瘤小鼠或应用于肿瘤球体。用多光子显微镜（MPM）可视化后，使用 Imaris 软件进行 T 细胞跟踪。Imaris 分析表明，与野生型 CTL 相比， CTL 在 PancOVA 肿瘤中的积累明显增加，肿瘤体积明显减小。与野生型（WT）CTL 相比， CTL 覆盖的距离（轨迹位移长度）更长，平均速度（平均轨迹速度）更高。瞬时速度分析表明， CTL 的阻滞轨迹（阻滞：<4 μm/min）的百分比更高。我们的数据表明，IL-18R 信号对 T 细胞效应子强度有贡献，值得进一步研究肿瘤内 T 细胞衰竭等现象。
