Chimeric antigen receptor T-cell（CAR-T）在血液肿瘤中尤为突出，但在实体肿瘤中并不突出，这主要是基于复杂的肿瘤免疫微环境。溶瘤病毒（OVs）是一种新兴的辅助治疗方法。OVs可以激活肿瘤病灶，诱导抗肿瘤免疫反应，从而增强CAR-T细胞的功能，可能提高治疗成功率。在本研究中，我们结合了针对碳酸酐酶9（CA9）的CAR-T细胞和携带趋化因子（C-C motif）配体5（CCL5）的溶瘤腺病毒（OAV），细胞因子白细胞介素-12（IL12），以探索这种组合策略的抗肿瘤效果。数据显示，Ad5-ZD55-hCCL5-hIL12能够感染和复制肾癌细胞系，并对裸鼠异种移植肿瘤产生适度的抑制作用。Ad5-ZD55-hCCL5-hIL12介导的IL12促进了CAR-T细胞中Stat4的磷酸化，并诱导CAR-T细胞分泌更多的干扰素-γ。我们还发现，Ad5-ZD55-hCCL5-hIL-12与CA9-CAR-T细胞的结合显著增加了CAR-T细胞在肿瘤中的浸润，延长了小鼠的存活时间，并抑制了免疫缺陷小鼠的肿瘤生长。Ad5-ZD55-mCCL5-mIL-12也能增加CD45+ CD3+ T细胞的浸润，延长免疫活性小鼠的存活时间。这些结果为溶瘤腺病毒和CAR-T细胞的组合提供了可行性，其展示了CAR-T治疗实体肿瘤的充分潜力和前景。版权所有 ©2023年作者。由Elsevier Ltd.出版发行。保留所有权利。

Chimeric antigen receptor T-cell (CAR-T) is particularly prominent in hematological but not in solid tumors, mainly based on the complex tumor immune microenvironment. Oncolytic virus (OVs) is an emerging adjuvant therapy method. OVs may prime tumor lesions to induce anti-tumor immune response, thereby enhancing CAR-T cells functionality and possibly increasing response rates. Here, we combined CAR-T cells targeting carbonic anhydrase 9 (CA9) and an oncolytic adenovirus (OAV) carrying chemokine (C-C motif) ligand 5 (CCL5), cytokine interleukin-12 (IL12) to explore the anti-tumor effects of this combination strategy. The data showed that Ad5-ZD55-hCCL5-hIL12 could infect and replicate in renal cancer cell lines and induced a moderate inhibition of xenografted tumor in nude mice. IL12 mediated by Ad5-ZD55-hCCL5-hIL12 promoted the phosphorylation of Stat4 in CAR-T cells, induced CAR-T cells to secrete more IFN-γ. We also found that Ad5-ZD55-hCCL5-hIL-12 combined with CA9-CAR-T cells significantly increased the infiltration of CAR-T cells in tumor mass, prolonged the survival of the mice and restrained tumor growth in immunodeficient mice. Ad5-ZD55-mCCL5-mIL-12 could also increase CD45+CD3+T cell infiltration and prolong mice survival in immunocompetent mice. These results provided feasibility for the combination of oncolytic adenovirus and CAR-T cells, which demonstrated the sufficient potential and prospects of CAR-T for the treatment of solid tumors.Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.