抗体介导的免疫原性表位递送将病毒特异性 CD8 T 细胞重定向到癌细胞是一种新兴且有前途的新治疗策略。这些所谓的抗体表位缀合物 (AEC) 依赖于靠近肿瘤表面的表位的蛋白水解释放，由 HLA I 类分子呈递，最终将病毒特异性 CD8 T 细胞重定向并激活至肿瘤细胞。我们将免疫原性 EBV-BRLF1 表位（前面有一个蛋白酶切割位点）融合到西妥昔单抗和曲妥珠单抗重链和/或轻链的 C 末端。我们评估了这些 AEC，发现尽管所有 AEC 都能够重定向 EBV 特异性 T 细胞，但与 AEC 相比，具有融合到重链 C 末端的表位的 AEC 导致 T 细胞激活水平更高具有与抗体轻链融合的相同表位。我们观察到所有 AEC 都取决于抗体靶点的存在，T 细胞激活水平与抗体靶点的表达水平相关，并且我们的 AEC 可以有效地将 BRLF1 表位传递到不同来源的癌细胞系中。乳腺癌、卵巢癌、肺癌和宫颈癌以及多发性骨髓瘤）。此外，在体内，AEC 有效降低了肿瘤负荷并提高了总生存期，与免疫检查点阻断相结合，可以进一步延长生存期。我们证明了这些基因融合的 AEC 在体外和体内将有效的 EBV 特异性 T 细胞重定向到癌症的潜力。© 2023。作者。

Antibody-mediated delivery of immunogenic epitopes to redirect virus-specific CD8+ T-cells towards cancer cells is an emerging and promising new therapeutic strategy. These so-called antibody-epitope conjugates (AECs) rely on the proteolytic release of the epitopes close to the tumor surface for presentation by HLA class I molecules to eventually redirect and activate virus-specific CD8+ T-cells towards tumor cells. We fused the immunogenic EBV-BRLF1 epitope preceded by a protease cleavage site to the C-terminus of the heavy and/or light chains of cetuximab and trastuzumab. We evaluated these AECs and found that, even though all AECs were able to redirect the EBV-specific T-cells, AECs with an epitope fused to the C-terminus of the heavy chain resulted in higher levels of T-cell activation compared to AECs with the same epitope fused to the light chain of an antibody. We observed that all AECs were depending on the presence of the antibody target, that the level of T-cell activation correlated with expression levels of the antibody target, and that our AECs could efficiently deliver the BRLF1 epitope to cancer cell lines from different origins (breast, ovarian, lung, and cervical cancer and a multiple myeloma). Moreover, in vivo, the AECs efficiently reduced tumor burden and increased the overall survival, which was prolonged even further in combination with immune checkpoint blockade. We demonstrate the potential of these genetically fused AECs to redirect the potent EBV-specific T-cells towards cancer in vitro and in vivo.© 2023. The Author(s).