Epstein-Barr virus (EBV) infections in healthy individuals are usually cleared by immune cells, wherein CD8⁺ T lymphocytes play the most important role. However, in some immunocompromised individuals, EBV infections can lead to the development of cancer in B, T, natural killer (NK) cells and epithelial cells. Most EBV-associated cancers express a limited number of virus-specific antigens such as latent membrane proteins (LMP1 and LMP2) and nuclear proteins (EBNA1, -2, EBNA3A, -B, -C, and EBNA-LP). These antigens represent true tumor-specific antigens and can be considered useful targets for T cell receptor (TCR) gene therapy to treat EBV-associated diseases. We used a TCR isolation platform based on a single major histocompatibility complex class I (MHC I) K562 cell library for the detection, isolation, and re-expression of TCRs targeting immunodominant peptide MHC (pMHC). Mature dendritic cells (mDCs) were pulsed with in vitro-transcribed (ivt) RNA encoding for the selected antigen to stimulate autologous T cells. The procedure allowed the mDCs to select an immunogenic epitope of the antigen for processing and presentation on the cell surface in combination with the most suitable MHC I molecule. We isolated eight EBV-specific TCRs. They recognize various pMHCs of EBV antigens LMP1, LMP2A, and EBNA3C, some of them described previously and some newly identified in this study. The TCR genes were molecularly cloned into retroviral vectors and the resultant TCR-engineered T cells secreted interferon-γ after antigen contact and were able to lyse tumor cells. The EBV-specific TCRs can be used as a basis for the generation of a TCR library, which provides a valuable source of TCRs for the production of EBV-specific T cells to treat EBV-associated diseases in patients with different MHC I types.

中文翻译：

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产生识别 LMP1、LMP2A 和 EBNA3C 免疫显性表位的 Epstein-Barr 病毒抗原特异性 T 细胞受体用于免疫治疗

健康个体中的 EB 病毒 (EBV) 感染通常被免疫细胞清除，其中 CD8 ⁺T淋巴细胞起着最重要的作用。然而，在一些免疫功能低下的个体中，EBV 感染可导致 B、T、自然杀伤 (NK) 细胞和上皮细胞发生癌症。大多数 EBV 相关癌症表达数量有限的病毒特异性抗原，例如潜伏膜蛋白（LMP1 和 LMP2）和核蛋白（EBNA1、-2、EBNA3A、-B、-C 和 EBNA-LP）。这些抗原代表真正的肿瘤特异性抗原，可以被认为是 T 细胞受体 (TCR) 基因治疗治疗 EBV 相关疾病的有用靶标。我们使用基于单个主要组织相容性复合物 I 类 (MHC I) K562 细胞库​​的 TCR 分离平台，用于检测、分离和重新表达靶向免疫显性肽 MHC (pMHC) 的 TCR。用体外脉冲成熟树突细胞 (mDC)-编码所选抗原的转录（ivt）RNA以刺激自体T细胞。该程序允许 mDCs 选择抗原的免疫原性表位，以便与最合适的 MHC I 分子组合在细胞表面进行加工和呈递。我们分离了八个 EBV 特异性 TCR。它们识别 EBV 抗原 LMP1、LMP2A 和 EBNA3C 的各种 pMHC，其中一些先前已描述，一些在本研究中新发现。将 TCR 基因分子克隆到逆转录病毒载体中，由此产生的 TCR 工程 T 细胞在抗原接触后分泌干扰素-γ，并能够裂解肿瘤细胞。EBV 特定的 TCR 可用作生成 TCR 库的基础，