EBV latent membrane protein 1 augments γδ T cell cytotoxicity against nasopharyngeal carcinoma by induction of butyrophilin molecules.
EBV latent membrane protein 1 augments γδ T cell cytotoxicity against nasopharyngeal carcinoma by induction of butyrophilin molecules.
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DOI：
10.7150/thno.78395
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发表时间：
2023
期刊：
影响因子：
12.4
通讯作者：
Cheung AKL
中科院分区：
文献类型：
--
作者：
Liu Y;Lui KS;Ye Z;Fung TY;Chen L;Sit PY;Leung CY;Mak NK;Wong KL;Lung HL;Tanaka Y;Cheung AKL
鼻咽癌 (NPC) 是一种多种癌症，没有明确的肿瘤抗原，与致癌的 Epstein-Barr 病毒 (EBV) 相关，通常诊断为晚期，生存率 <40%。目前的放疗和化疗效果低下且产生不良反应，需要新的治疗方法。对此，利用γδ T细胞的过继性免疫疗法具有潜力，但需要与嗜丁蛋白2A1和3A1蛋白表达相结合才能达到杀瘤效果。方法：扩增人 γδ T 细胞（用 Zol 或 PTA）并用于针对用 EBV EBNA1 靶向肽 (L2)P4 处理的 NPC 细胞进行细胞毒性测定。通过流式细胞术和Western blot检测(L2)P4对NPC细胞中BTN2A1/BTN3A1表达的影响。建立了携带 NPC 的 NSG 小鼠模型来测试 P4 和过继性 γδ T 细胞的有效性。对鼻咽癌组织切片进行免疫荧光检查 γδ T 细胞的存在以及 BTN2A1 和 BTN3A1 的表达。通过 qRT-PCR 评估 (L2)P4 处理后的 EBV 基因表达，并通过转染、报告基因测定、Western blot 和抑制实验检查 LMP1、NLRC5 和 BTN2A1/BTN3A1 的关系。结果：Zol 或 PTA 扩增了 γδ T 细胞的 Vδ2 子集，对某些 NPC 细胞发挥杀伤作用。 (L2)P4 重新激活潜在的 EBV，从而增加 BTN2A1 和 BTN3A1 的表达，并赋予 Vδ2 T 细胞体外细胞毒性更高的敏感性，并通过 Vδ2 T 细胞的过继转移增强体内肿瘤消退。从机制上讲，(L2)P4 诱导 EBV LMP1，导致 IFN-γ/p-JNK 和 NLRC5 激活，随后刺激 BTN2A1 和 BTN3A1 的表达。结论：本研究证明了使用 EBV 靶向探针 (L2)P4 和过继性 γδ T 细胞作为一种有前景的鼻咽癌组合免疫疗法的有效性。 LMP1-IFN-γ/p-JNK-NLRC5-BTN2A1/BTN3A1 轴的鉴定可能会带来针对 NPC 和其他 EBV+ 肿瘤的新见解和治疗靶点。
Nasopharyngeal carcinoma (NPC) is a diverse cancer with no well-defined tumor antigen, associated with oncogenic Epstein-Barr Virus (EBV), and with usually late-stage diagnosis and survival <40%. Current radiotherapy and chemotherapy have low effectiveness and cause adverse effects, which calls for the need of new therapy. In this regard, adoptive immunotherapy using γδ T cells has potential, but needs to be coupled with butyrophilin 2A1 and 3A1 protein expression to achieve tumoricidal effect. Methods: Human γδ T cells were expanded (with Zol or PTA) and used for cytotoxicity assay against NPC cells, which were treated with the EBV EBNA1-targeting peptide (L2)P4. Effect of (L2)P4 on BTN2A1/BTN3A1 expression in NPC cells was examined by flow cytometry and Western blot. An NPC-bearing NSG mice model was established to test the effectiveness of P4 and adoptive γδ T cells. Immunofluorescence was performed on NPC tissue sections to examine the presence of γδ T cells and expression of BTN2A1 and BTN3A1. EBV gene expression post-(L2)P4 treatment was assessed by qRT-PCR, and the relationship of LMP1, NLRC5 and BTN2A1/BTN3A1 was examined by transfection, reporter assay, Western blot, and inhibition experiments. Results: Zol- or PTA-expanded the Vδ2 subset of γδ T cells that exerted killing against certain NPC cells. (L2)P4 reactivates latent EBV, which increased BTN2A1 and BTN3A1 expression and conferred higher susceptibility towards Vδ2 T cells cytotoxicity in vitro, as well as enhanced tumor regression in vivo by adoptive transfer of Vδ2 T cells. Mechanistically, (L2)P4 induced EBV LMP1, leading to IFN-γ/p-JNK and NLRC5 activation, and subsequently stimulated the expression of BTN2A1 and BTN3A1. Conclusions: This study demonstrated the effectiveness of using the EBV-targeting probe (L2)P4 and adoptive γδ T cells as a promising combinatorial immunotherapy against NPC. The identification of the LMP1-IFN-γ/p-JNK-NLRC5-BTN2A1/BTN3A1 axis may lead to new insight and therapeutic targets against NPC and other EBV+ tumors.
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影响因子：
16.6
作者：
Liu Y;He S;Wang XL;Peng W;Chen QY;Chi DM;Chen JR;Han BW;Lin GW;Li YQ;Wang QY;Peng RJ;Wei PP;Guo X;Li B;Xia X;Mai HQ;Hu XD;Zhang Z;Zeng YX;Bei JX
通讯作者：
Bei JX
影响因子：
11.4
作者：
Gires, O;Kohlhuber, F;Hammerschmidt, W
通讯作者：
Hammerschmidt, W
影响因子：
7.5
作者：
Cheung, Allen Ka Loon;Huang, Yiru;Chen, Zhiwei
通讯作者：
Chen, Zhiwei
影响因子：
4.5
作者：
Abdulamir, A. S.;Hafidh, R. R.;Abdulmuhaimen, N.;Abubakar, F.;Abbas, K. A.
通讯作者：
Abbas, K. A.
影响因子：
5.4
作者：
Bentz, Gretchen L.;Whitehurst, Christopher B.;Pagano, Joseph S.
通讯作者：
Pagano, Joseph S.