肺癌仍属于难以治疗的恶性肿瘤之一，是全球癌症相关死亡的主要原因。靶向治疗和检查点抑制剂的引入改善了治疗效果，然而，大多数晚期非小细胞肺癌（NSCLC）患者最终仍对这些治疗方法失效。因此，检查点难治性/耐药性NSCLC存在重大临床需求。本研究在免疫活性的小鼠NSCLC模型中测试了aPD-1和携带TNFα和IL-2的腺病毒（Ad5-CMV-mTNFα/mIL-2）的联合应用。此外，尽管局部给药已成为病毒治疗的标准，但本研究采用静脉注射的方式进行治疗，以便促进临床转化和日常应用。我们表明，联合应用aPD-1和静脉注射携带武装病毒的腺病毒可显著抑制肿瘤生长，即使存在中和抗体。我们观察到细胞毒性的肿瘤组织浸润淋巴细胞的频率增加，包括肿瘤特异性细胞。联合治疗导致免疫抑制性肿瘤相关巨噬细胞的百分比降低，并改善树突状细胞的成熟。此外，我们观察到接受aPD-1和病毒联合治疗的组别在二级淋巴器官中展开了肿瘤特异性记忆T细胞组分。然而，尽管非复制性的Ad5-CMV-mTNFα/mIL-2病毒在小鼠模型中具有高水平的转基因表达，但它并不完全反映人类的临床结果。因此，通过在完全适应TNFα和IL-2武装的溶瘤腺病毒TILT-123的NSCLC离体模型中补充我们的发现。总的来说，我们的数据证明了通过静脉注射携带TNFα和IL-2的腺病毒能够增强aPD-1的抗肿瘤效果，并值得进一步在临床试验中进行深入研究。© 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.

Lung cancer remains among the most difficult-to-treat malignancies and is the leading cause of cancer-related deaths worldwide. The introduction of targeted therapies and checkpoint inhibitors has improved treatment outcomes; however, most patients with advanced-stage non-small cell lung cancer (NSCLC) eventually fail these therapies. Therefore, there is a major unmet clinical need for checkpoint refractory/resistant NSCLC. Here, we tested the combination of aPD-1 and adenovirus armed with TNFα and IL-2 (Ad5-CMV-mTNFα/mIL-2) in an immunocompetent murine NSCLC model. Moreover, although local delivery has been standard for virotherapy, treatment was administered intravenously to facilitate clinical translation and putative routine use. We showed that treatment of tumor-bearing animals with aPD-1 in combination with intravenously injected armed adenovirus significantly decreased cancer growth, even in the presence of neutralizing antibodies. We observed an increased frequency of cytotoxic tumor-infiltrating lymphocytes, including tumor-specific cells. Combination treatment led to a decreased percentage of immunosuppressive tumor-associated macrophages and an improvement in dendritic cell maturation. Moreover, we observed expansion of the tumor-specific memory T cell compartment in secondary lymphoid organs in the group that received aPD-1 with the virus. However, although the non-replicative Ad5-CMV-mTNFα/mIL-2 virus allows high transgene expression in the murine model, it does not fully reflect the clinical outcome in humans. Thus, we complemented our findings using NSCLC ex vivo models fully permissive for the TNFα and IL-2- armed oncolytic adenovirus TILT-123. Overall, our data demonstrate the ability of systemically administered adenovirus armed with TNFα and IL-2 to potentiate the anti-tumor efficacy of aPD-1 and warrant further investigation in clinical trials.© 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.