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

We demonstrated PD-L1 expression in <i>EGFR</i> and<i> KRAS </i>mutant and <i>EGFR/KRAS-</i>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 <i>EGFR</i>-MT, <i>KRAS</i>-MT<i>E</i><i>G</i><i>FR/KRAS</i>-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 <i>EGFR</i>-MT, <i>KRAS</i>-MT and <i>EGFR/KRAS</i>-WT NSCLC tumor organoids was detected by flow cytometry.本研究通过干扰素-γ（IFN-γ）处理或共培养的方式，以未处理的肿瘤类器官为对照，检测了表皮生长因子受体（EGFR）突变型、Kirsten大鼠肉瘤病毒癌基因同源物（KRAS）突变型及EGFR/KRAS双野生型肿瘤类器官的程序性死亡受体配体1（PD-L1）表达水平。经抗PD-1（程序性死亡受体1）治疗后，本研究通过流式细胞术检测了PD-1对T细胞的调控作用，并通过检测CD8阳性（CD8⁺）T细胞中CD107a、穿孔素（perforin）与颗粒酶B（Granzyme B）的持续表达水平，评估效应T细胞的细胞毒性功能。此外，本研究通过酶联免疫吸附试验（ELISA）分别检测了非小细胞肺癌（NSCLC）患者与非癌个体的白细胞介素17（IL-17）浓度，以及NSCLC患者血浆中的IL-17水平。在肿瘤类器官-T细胞共培养体系中，本研究探究了EGFR突变型、KRAS突变型及EGFR/KRAS双野生型肿瘤类器官上IL-17与白细胞介素17受体（IL-17R）的共表达情况；同时通过流式细胞术检测了CD4阳性（CD4⁺）与CD8阳性（CD8⁺）T细胞表面的IL-17R表达水平。最后，本研究通过流式细胞术检测了IL-17阻断治疗对EGFR突变型、KRAS突变型及EGFR/KRAS双野生型非小细胞肺癌肿瘤类器官中PD-1阻断治疗疗效的调控作用。