免疫原性细胞死亡（ICD）已成为癌症免疫治疗的一种新选择。 ICD 的关键决定因素包括抗原性（抗原的存在）和佐剂性，其中涉及损伤相关分子模式 (DAMP) 以及各种细胞因子和趋化因子的释放。 CX3CL1，也称为神经趋化素或 fractalkine，是一种参与细胞信号传导和免疫细胞相互作用的趋化因子。 CX3CL1 被称为“找到我”信号，可刺激免疫细胞向垂死细胞趋化，促进胞吞作用和抗原呈递。然而，在 ICD 背景下，尚不确定 CX3CL1 是否是 ICD 效应的重要介质。在本研究中，我们研究了 CX3CL1 在米托蒽醌 (MTX) 诱导的癌细胞免疫原性细胞凋亡中的复杂作用。 Luminex xMAP 技术用于量化小鼠细胞因子、趋化因子和生长因子，以确定经历 ICD 的小鼠纤维肉瘤 MCA205 和黑色素瘤 B16-F10 细胞释放的关键调节细胞因子。此外，采用小鼠肿瘤预防性疫苗接种模型来分析 CX3CL1 对 ICD 中针对 MCA205 细胞的适应性免疫反应激活的影响。此外，对 98 名黑色素瘤患者的 TCGA-SKCM 公共数据集的彻底分析揭示了 CX3CL1 及其受体 CX3CR1 在黑色素瘤患者中的作用。我们的研究结果表明，凋亡的 MCA205 和 B16-F10 细胞（无论细胞类型如何）中 CX3CL1 的释放增强，但如果它们正在经历铁死亡或意外坏死，则不会。此外，在小鼠预防性肿瘤疫苗接种模型中，将重组CX3CL1添加到非免疫原剂量的MTX处理的​​凋亡性死亡癌细胞中，诱导了强烈的免疫原性反应，有效提高了小鼠的存活率。此外，对黑色素瘤患者数据的分析显示，表达 CX3CR1 的 CD8 T 细胞水平升高的个体的生存率有所提高。这些数据共同强调了 CX3CL1 的释放在引发针对垂死癌细胞的免疫原性反应中的重要性，并表明 CX3CL1 可能作为赋予细胞凋亡免疫原性的关键开关。版权所有 © 2024 Naessens、Demuynck、Vershinina、Efimova、Saviuk、De Smet、Mishchenko、Vedunova、Krysko、Catanzaro 和 Krysko。

Immunogenic cell death (ICD) has emerged as a novel option for cancer immunotherapy. The key determinants of ICD encompass antigenicity (the presence of antigens) and adjuvanticity, which involves the release of damage-associated molecular patterns (DAMPs) and various cytokines and chemokines. CX3CL1, also known as neurotactin or fractalkine, is a chemokine involved in cellular signalling and immune cell interactions. CX3CL1 has been denoted as a "find me" signal that stimulates chemotaxis of immune cells towards dying cells, facilitating efferocytosis and antigen presentation. However, in the context of ICD, it is uncertain whether CX3CL1 is an important mediator of the effects of ICD.In this study, we investigated the intricate role of CX3CL1 in immunogenic apoptosis induced by mitoxantrone (MTX) in cancer cells. The Luminex xMAP technology was used to quantify murine cytokines, chemokines and growth factors to identify pivotal regulatory cytokines released by murine fibrosarcoma MCA205 and melanoma B16-F10 cells undergoing ICD. Moreover, a murine tumour prophylactic vaccination model was employed to analyse the effect of CX3CL1 on the activation of an adaptive immune response against MCA205 cells undergoing ICD. Furthermore, thorough analysis of the TCGA-SKCM public dataset from 98 melanoma patients revealed the role of CX3CL1 and its receptor CX3CR1 in melanoma patients.Our findings demonstrate enhanced CX3CL1 release from apoptotic MCA205 and B16-F10 cells (regardless of the cell type) but not if they are undergoing ferroptosis or accidental necrosis. Moreover, the addition of recombinant CX3CL1 to non-immunogenic doses of MTX-treated, apoptotically dying cancer cells in the murine prophylactic tumour vaccination model induced a robust immunogenic response, effectively increasing the survival of the mice. Furthermore, analysis of melanoma patient data revealed enhanced survival rates in individuals exhibiting elevated levels of CD8+ T cells expressing CX3CR1.These data collectively underscore the importance of the release of CX3CL1 in eliciting an immunogenic response against dying cancer cells and suggest that CX3CL1 may serve as a key switch in conferring immunogenicity to apoptosis.Copyright © 2024 Naessens, Demuynck, Vershinina, Efimova, Saviuk, De Smet, Mishchenko, Vedunova, Krysko, Catanzaro and Krysko.