Retargeting NK92 cells using an HLA-A2-restricted, EBNA3C-specific chimeric antigen receptor
Retargeting NK92 cells using an HLA-A2-restricted, EBNA3C-specific chimeric antigen receptor
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DOI：
10.1038/cgt.2011.66
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发表时间：
2012-02-01
影响因子：
6.4
通讯作者：
Cheung, N-K V.
中科院分区：
文献类型：
--
作者：
Tassev, D. V.;Cheng, M.;Cheung, N-K V.
过继性细胞免疫疗法的进展为癌症患者带来了几种有希望的选择。通过在重组人白细胞抗原（HLA）-A2-肽复合物上淘选，从人噬菌体展示文库中分离单链可变区片段（scFv）。对HLA-A2特异性的scFv（EBNA克隆315）进行了充分表征，所述scFv携带来源于EB病毒潜伏蛋白EBNA 3C的10个氨基酸的肽（LLDFVRFMGV）。EBNA克隆315对其靶向的T细胞表位（TCE）显示出精确的特异性，并且不与携带无关肽的游离肽、HLA-A2复合物或HLA-A2（-）细胞交叉反应。此外，在工程改造成scFv-Fc融合蛋白后，我们能够确定其亲和力、检测灵敏度和诱导抗体依赖性细胞毒性（ADCC）的能力。作为原理证明，EBNA克隆315的嵌合抗原受体（CAR）形式用于重编程NK 92 MI细胞。表达CAR的NK 92 MI细胞对靶向TCE显示出高度特异性和有效的细胞毒性，检测灵敏度约为25个分子，溶细胞能力是scFv-Fc介导的ADCC的三倍。这是第一次，我们使用CAR成功地将非T细胞重编程为特定的TCE。Cancer Gene Therapy（2012）19，84-100; doi：10.1038/cgt.2011.66; 2011年10月7日在线发表
Advances in adoptive cell immunotherapy have led to several promising options for cancer patients. Single-chain variable fragments (scFvs) were isolated from a human phage display library by panning on recombinant human leukocyte antigen (HLA)-A2-peptide complexes. A scFv (EBNA Clone 315) specific for HLA-A2 carrying a 10 amino acid peptide (LLDFVRFMGV) derived from the Epstein-Barr virus latent protein EBNA3C was fully characterized. EBNA Clone 315 displayed exquisite specificity toward its targeted T-cell epitope (TCE) and did not cross-react with the free peptide, HLA-A2 complexes, which carried irrelevant peptides, or HLA-A2(-) cells. Furthermore, after engineering into a scFv-Fc fusion protein, we were able to determine its affinity, detection sensitivity, and ability to induce antibody-dependent cellular cytotoxicity (ADCC). As a proof-of-principle, a chimeric antigen receptor (CAR) version of EBNA Clone 315 was used to reprogram NK92MI cells. CAR-expressing NK92MI cells showed highly specific and potent cytotoxicity toward the targeted TCE, with detection sensitivity of approximately 25 molecules and cytolytic capacity threefold greater than scFv-Fc-mediated ADCC. For the first time, we show the successful reprogramming of non-T cells toward a specific TCE using a CAR. Cancer Gene Therapy (2012) 19, 84-100; doi: 10.1038/cgt.2011.66; published online 7 October 2011