HLA-A2.1 限制性 ECM1 衍生表位 LA 通过 DC 交叉激活启动 CD8 T 和 NK 细胞：一种新型治疗性肿瘤疫苗。
HLA-A2.1-restricted ECM1-derived epitope LA through DC cross-activation priming CD8 T and NK cells: a novel therapeutic tumour vaccine.
机构信息
Department of Pharmacology, School of Pharmacy, China Medical University, No. 13, Beihai Road, Dadong District, Shenyang, Liaoning Province, China.
Liaoning Key Laboratory of Molecular Targeted Antitumour Drug Development and Evaluation, Liaoning Cancer Immune Peptide Drug Engineering Technology Research Centre, Key Laboratory of Precision Diagnosis and Treatment of Gastrointestinal Tumours, Ministry of Education, China Medical University, No.77, Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province, China.
出版信息
J Hematol Oncol. 2021 Apr 28;14(1):71. doi: 10.1186/s13045-021-01081-7.
BACKGROUND
CD8 T cell-mediated adaptive cellular immunity and natural killer (NK) cell-mediated innate immunity both play important roles in tumour immunity. This study aimed to develop therapeutic tumour vaccines based on double-activation of CD8 T and NK cells.
METHODS
The immune Epitope database, Molecular Operating Environment software, and enzyme-linked immunosorbent assay were used for epitope identification. Flow cytometry, confocal microscopy, UPLC-QTOF-MS, and RNA-seq were utilized for evaluating immunity of PBMC-derived DCs, CD8 T or NK cells and related pathways. HLA-A2.1 transgenic mice combined with immunologically reconstituted tumour-bearing mice were used to examine the antitumour effect and safety of epitope vaccines.
RESULTS
We identified novel HLA-A2.1-restricted extracellular matrix protein 1(ECM1)-derived immunodominant epitopes in which LA induced a potent immune response. We also found that LA-loaded DCs upregulated the frequency of CD3/CD8 T cells, CD45RO/CD69 activated memory T cells, and CD3/CD16/CD56 NK cells. We demonstrated cytotoxic granule release of LA/DC-CTLs or LA/DC-NK cells and cytotoxicity against tumour cells and microtissue blocks via the predominant IFN-γ/perforin/granzyme B cell death pathway. Further investigating the mechanism of LA-mediated CD8 T activation, we found that LA could be internalized into DCs through phagocytosis and then formed a LA-MHC-I complex presented onto the DC surface for recognition of the T cell receptor to upregulate Zap70 phosphorylation levels to further activate CD8 T cells by DC-CTL interactions. In addition, LA-mediated DC-NK crosstalk through stimulation of the TLR4-p38 MAPK pathway increased MICA/B expression on DCs to interact with NKG2D for NK activation. Promisingly, LA could activate CD8 T cells and NK cells simultaneously via interacting with DCs to suppress tumours in vivo. Moreover, the safety of LA was confirmed.
CONCLUSIONS
LA-induced immune antitumour activity through DC cross-activation with CD8 T and NK cells, which demonstrated proof-of-concept evidence for the capability and safety of a novel therapeutic tumour vaccine.
背景
CD8 T 细胞介导的适应性细胞免疫和自然杀伤（NK）细胞介导的固有免疫在肿瘤免疫中都发挥着重要作用。本研究旨在开发基于双重激活 CD8 T 和 NK 细胞的治疗性肿瘤疫苗。
方法
使用免疫表位数据库、分子操作环境软件和酶联免疫吸附试验进行表位鉴定。流式细胞术、共聚焦显微镜、UPLC-QTOF-MS 和 RNA-seq 用于评估 PBMC 来源的 DC、CD8 T 或 NK 细胞的免疫和相关途径。利用 HLA-A2.1 转基因小鼠结合免疫重建荷瘤小鼠来检验表位疫苗的抗肿瘤作用和安全性。
结果
我们鉴定了新型 HLA-A2.1 限制性细胞外基质蛋白 1（ECM1）衍生的免疫显性表位，其中 LA 诱导了强烈的免疫反应。我们还发现，LA 负载的 DC 上调了 CD3/CD8 T 细胞、CD45RO/CD69 激活的记忆 T 细胞和 CD3/CD16/CD56 NK 细胞的频率。我们证明了 LA/DC-CTL 或 LA/DC-NK 细胞的细胞毒性颗粒释放以及通过主要的 IFN-γ/穿孔素/颗粒酶 B 细胞死亡途径对肿瘤细胞和微组织块的细胞毒性。进一步研究 LA 介导的 CD8 T 激活的机制，我们发现 LA 可以通过吞噬作用内化到 DC 中，然后形成呈现在 DC 表面的 LA-MHC-I 复合物，以识别 T 细胞受体来上调 Zap70 磷酸化水平，从而通过 DC-CTL 相互作用进一步激活 CD8 T 细胞。此外，LA 通过刺激 TLR4-p38 MAPK 途径介导的 DC-NK 串扰增加了 DC 上的 MICA/B 表达，以与 NKG2D 相互作用激活 NK。有希望的是，LA 可以通过与 DC 相互作用来同时激活 CD8 T 细胞和 NK 细胞，从而在体内抑制肿瘤。此外，还证实了 LA 的安全性。
结论
LA 通过与 DC 交叉激活 CD8 T 和 NK 细胞诱导免疫抗肿瘤活性，为新型治疗性肿瘤疫苗的有效性和安全性提供了概念验证证据。
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