Supporting data for "Modulation of IL-17 Enhances the Efficiency of Anti-PD-1 Therapy in Non-small-cell Lung Cancer"

We demonstrated PD-L1 expression in EGFR and KRAS mutant and EGFR/KRAS-wildtype tumor organoids by IFN-γ or co-culture compared with tumor organoids. PD-1 action on T cells and the cytotoxicity of effector T cells through sustained CD107a, perforin and Granzyme B expression in CD8+ T cells were detected by flow cytometry after anti-PD-1 treatment. On the other hand, IL-17 concentration in NSCLC patients and non-cancer individuals was detected by ELISA. The level of IL-17 in NSCLC patients' plasma was detected by ELISA. In the tumor organoids-T cells co-culture system, we investigated the concomitant presence of IL-17 and IL-17R expression on EGFR-MT, KRAS-MTEGFR/KRAS-WT tumor organoids. IL-17R expression on CD4+ and CD8+ T cells was detected by flow cytometry. Modulation of IL-17 blockade on the efficacy of PD-1 blockade in EGFR-MT, KRAS-MT and EGFR/KRAS-WT NSCLC tumor organoids was detected by flow cytometry.

本研究通过IFN-γ或共培养方法，在EGFR和KRAS突变体以及EGFR/KRAS野生型肿瘤类器官中，展示了PD-L1的表达水平。在抗PD-1治疗后，通过流式细胞术检测到T细胞的PD-1作用以及效应T细胞的细胞毒性，表现为CD8+ T细胞中持续表达的CD107a、穿孔素和颗粒酶B。另一方面，通过ELISA技术检测了非小细胞肺癌患者和非癌症个体体内的IL-17浓度，以及在肺癌患者血浆中的IL-17水平。在肿瘤类器官与T细胞的共培养体系中，本研究探讨了IL-17及其受体在EGFR-MT、KRAS-MTEGFR/KRAS-WT肿瘤类器官中的共存表达。通过流式细胞术检测了CD4+和CD8+ T细胞上的IL-17R表达。此外，还通过流式细胞术检测了IL-17阻断对PD-1阻断在EGFR-MT、KRAS-MT和EGFR/KRAS-WT非小细胞肺癌肿瘤类器官疗效的影响。