尽管肿瘤浸润淋巴细胞（TILs）的存在与卵巢癌（OvCa）患者的生存预后改善有关，但TIL疗法的益处有限。在本研究中，我们评估了编码人类变异白细胞介素-2（vIL-2）细胞因子的溶瘤腺病毒Ad5 / 3-E2F-d24-vIL2（vIL-2病毒），也被称为TILT-452，作为一种免疫治疗策略，以增强对晚期OvCa肿瘤的TIL反应性。切除的人类OvCa肿瘤组织被处理成单细胞悬液，并扩增自体的TILs。OvCa肿瘤标本与TILs加vIL-2病毒进行共培养，并通过细胞阻抗测量实时评估细胞杀伤能力。组合疗法进一步通过患者源性异种移植（PDX）卵巢癌小鼠模型进行评估。与TILs单独治疗相比，vIL-2病毒加TILs的组合疗法在体外对癌细胞的杀伤效果最佳。这些结果得到了体内实验的支持，当将vIL-2病毒加入TIL疗法时，获得了最佳的OvCa肿瘤控制效果。此外，所提出的治疗方案诱导了高度细胞毒性表型的形成，表现为治疗肿瘤中NK细胞、CD4+ T细胞和CD8+ T细胞的粒酶B强度增加。我们的结果表明，Ad5 / 3-E2F-d24-vIL2疗法在治疗人类OvCa肿瘤时始终能够改善TIL疗法的细胞毒性。©2023.作者。

While the presence of tumor-infiltrating lymphocytes (TILs) associates with improved survival prognosis in ovarian cancer (OvCa) patients, TIL therapy benefit is limited. Here, we evaluated an oncolytic adenovirus coding for a human variant IL-2 (vIL-2) cytokine, Ad5/3-E2F-d24-vIL2 (vIL-2 virus), also known as TILT-452, as an immunotherapeutic strategy to enhance TIL responsiveness towards advanced stage OvCa tumors. Fragments of resected human OvCa tumors were processed into single-cell suspensions, and autologous TILs were expanded from said samples. OvCa tumor specimens were co-cultured with TILs plus vIL-2 virus, and cell killing was assessed in real time through cell impedance measurement. Combination therapy was further evaluated in vivo through a patient-derived xenograft (PDX) ovarian cancer murine model. The combination of vIL-2 virus plus TILs had best cancer cell killing ex vivo compared to TILs monotherapy. These results were supported by an in vivo experiment, where the best OvCa tumor control was obtained when vIL-2 virus was added to TIL therapy. Furthermore, the proposed therapy induced a highly cytotoxic phenotype demonstrated by increased granzyme B intensity in NK cells, CD4+ T, and CD8+ T cells in treated tumors. Our results demonstrate that Ad5/3-E2F-d24-vIL2 therapy consistently improved TILs therapy cytotoxicity in treated human OvCa tumors.© 2023. The Author(s).