共表达IL-7/CCL19的新抗原反应性T细胞对小鼠肺癌的抗肿瘤研究  

Antitumor Study of Neoantigen-reactive T Cells Co-expressing IL-7 and CCL19 in Mouse Lung Cancer

作　　者：吴迪[1] 李晨辉 汪艳 何正强[1] 金常娥[1] 郭敏 陈荣昌 周承志[2] Di WU;Chenhui LI;Yan WANG;Zhengqiang HE;Chang’e JIN;Min GUO;Rongchang CHEN;Chengzhi ZHOU(Department of Pulmonary and Critical Care Medicine,Shenzhen Institute of Respiratory Diseases,The Second Clinical Medical Col-lege,Jinan University(Shenzhen People’s Hospital),The First Affiliated Hospital of Southern University of Science and Technology(Shenzhen People’s Hospital),Shenzhen 518020,China;Pulmonary and Critical Care Medicine,Guangzhou Institute of Respiratory Health,National Clinical Research Center for Respiratory Disease,National Center for Respiratory Medicine,State Key Laboratory of Respiratory Diseases,The First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China)

机构地区：[1]暨南大学第二临床医学院(深圳市人民医院)深圳市呼吸疾病研究所肺重症医学科,南方科技大学第一附属医院(深圳市人民医院),深圳518020 [2]广州医科大学第一附属医院肺与重症医学研究所,国家呼吸疾病临床研究中心,国家呼吸医学中心,呼吸疾病国家重点实验室,广州510120

出　　处：《中国肺癌杂志》2024年第7期504-513,共10页Chinese Journal of Lung Cancer

基　　金：广东省医学科学技术研究基金项目(No.A2020487);呼吸疾病国家重点实验室开放课题研究项目(No.SKLRD-OP-202011)资助。

摘　　要：背景与目的新抗原反应性T细胞(neoantigen reactive T cell,NRT)具有抑制表达特异性新抗原的肿瘤生长的能力。然而,由于免疫浸润困难和肿瘤微环境的抑制,NRT在实体瘤中的治疗效果有限。本研究针对小鼠肺癌细胞设计出可以同时表达白细胞介素7(interleukin 7,IL-7)和趋化因子19(chemokine C-C motif ligand 19,CCL19)的NRT细胞(7×19 NRT),并对7×19 NRT细胞和常规NRT细胞的抗肿瘤效果差异进行了评估。方法针对小鼠Lewis肺癌细胞(Lewis lung carcinoma,LLC)进行了新一代测序和新抗原预测,制备了RNA疫苗,培养了NRT细胞,构建了编码IL-7和CCL19的逆转录病毒载体,转导NRT细胞并成功表达IL-7和CCL19,成功获得了7×19 NRT,并在小鼠体内外对其抗肿瘤效果进行了评估。结果7×19 NRT细胞通过分泌IL-7和CCL19显著增强T细胞的增殖和侵袭能力,在小鼠肺癌中实现了显著的抑瘤作用,延长了小鼠的生存期。经7×19 NRT治疗后,T细胞浸润肿瘤组织及肿瘤组织坏死显著增加。此外,7×19 NRT治疗与常规NRT治疗均安全。结论通过IL-7和CCL19的表达,NRT细胞的抗实体瘤能力显著增强,这是一种对NRT安全有效的基因修饰。Background and objective Neoantigen reactive T cell(NRT)has the ability to inhibit the growth of tumors expressing specific neoantigens.However,due to the difficult immune infiltration and the inhibition of tumor microen-vironment,the therapeutic effect of NRT in solid tumors is limited.In this study,we designed NRT cells(7×19 NRT)that can express both interleukin-7(IL-7)and chemokine C-C motif ligand 19(CCL19)in mouse lung cancer cells,and evaluated the difference in anti-tumor effect between 7×19 NRT cells and conventional NRT cells.Methods We performed next-generation sequencing and neoantigen prediction for mouse Lewis lung carcinoma(LLC),prepared RNA vaccine,cultured NRT cells,constructed retroviral vectors encoding IL-7 and CCL19,transduced NRT cells and IL-7 and CCL19 were successfully ex-pressed,and 7×19 NRT was successfully obtained.The anti-tumor effect was evaluated in vivo and in vitro in mice.Results The 7×19 NRT cells significantly enhanced the proliferation and invasion ability of T cells by secreting IL-7 and CCL19,achieved significant tumor inhibition in the mouse lung cancer and extended the survival period of mice.The T cell infiltration into tumor tissue and the necrosis of tumor tissue increased significantly after 7×19 NRT treatment.In addition,both 7×19 NRT treatment and conventional NRT treatment were safe.Conclusion The anti-solid tumor ability of NRT cells is significantly enhanced by the arming of IL-7 and CCL19,which is a safe and effective genetic modification of NRT.

关 键 词：新抗原 白细胞介素7 趋化因子19 肺肿瘤 新抗原反应性T细胞 

分 类 号：R734.2[医药卫生—肿瘤]