辐照（IR）能够引起肿瘤中的免疫原性细胞死亡（ICD），但很少导致远隔效应（AE）。然而，将IR与免疫检查点抑制剂相结合已经显示出通过诱导AE来引发 anecdotes级的成功。在本研究中，我们旨在通过利用抗酗酒药物二硫化四氨和铜配合物（DSF / Cu）诱导肿瘤ICD来增强IR诱导的免疫应答，并产生可重复的AEs。我们在体内外测量了ICD。在小鼠肿瘤模型中，我们通过肿瘤内注射DSF / Cu，紧接着局部肿瘤IR，创建了一种体内癌疫苗。我们通过原发肿瘤的排斥来确定抗癌应答，并通过肿瘤再挑战和AEs（即自发性肺转移）来评估全身免疫应答。此外，我们分析了免疫细胞亚群，并在接种小鼠的肿瘤微环境（TME）和血液中定量了促炎和免疫抑制性趋化因子/细胞因子。通过调查免疫细胞消耗对疫苗诱导的抗癌应答的影响。结果显示，DSF / Cu和IR在低氧环境下诱导的ICD比在正氧环境下更有效。 DSF / Cu和IR的低剂量肿瘤内注射在小鼠模型中显示了强大的原发和再挑战肿瘤的抗癌效果和可观的AEs。这些疫苗还增加了4T1模型中的CD8+和CD4 +细胞数量，同时减少了Tregs和骨髓源性抑制细胞，并增加了MCa-M3C模型中的CD8 +，DC细胞数量，并减少了Treg细胞数量。去除CD8+和CD4+细胞会消除疫苗的抗癌应答。此外，接种肿瘤携带小鼠表现出增加的TNFα水平和降低的免疫抑制趋化因子/细胞因子水平。总之，我们的新方法产生了抗癌免疫应答，导致再挑战后肿瘤发病缺失或较低，以及强大的AEs，即肿瘤携带小鼠中的自发性肺转移的缺失或减少。这种方法易于在临床环境中推广使用，并可能增加肿瘤患者中IR引起的AEs。

Irradiation (IR) induces immunogenic cell death (ICD) in tumors, but it rarely leads to the abscopal effect (AE). However, combining IR with immune checkpoint inhibitors has shown anecdotal success in inducing AEs. In this study, we aimed to enhance the IR-induced immune response and generate reproducible AEs using the anti-alcoholism drug disulfiram (DSF) and copper complex (DSF/Cu) via induction of tumor ICD. We measured ICD in vitro and in vivo. In mouse tumor models, DSF/Cu was injected intratumorally followed by localized tumor IR, creating an in situ cancer vaccine. We determined the anti-cancer response by primary tumor rejection and assessed systemic immune responses by tumor rechallenge and the occurrence of AEs, i.e., spontaneous lung metastasis. Additionally, we analyzed immune cell subsets and quantified proinflammatory and immunosuppressive chemokines/cytokines in the tumor microenvironment (TME) and blood of the vaccinated mice. Immune cell depletion was investigated for its effects on the vaccine-induced anti-cancer response. The results showed that DSF/Cu and IR induced more potent ICD under hypoxia than normoxia in vitro. Low-dose intratumoral injection of DSF/Cu and IR demonstrated strong anti-primary and -rechallenged tumor effects and robust AEs in mouse models. These vaccinations also increased CD8 + and CD4 + cell numbers while decreasing Tregs and myeloid-derived suppressor cells in the 4T1 model, and increased CD8+, DC, and decreased Treg cell numbers in the MCa-M3C model. Depleting both CD8 + and CD4 + cells abolished the vaccine's anti-cancer response. Moreover, vaccinated tumor-bearing mice exhibited increased TNFα levels and reduced levels of immunosuppressive chemokines/cytokines. In conclusion, our novel approach generated an anti-cancer immune response, resulting in a lack of or low tumor incidence post-rechallenge and robust AEs, i.e., the absence of or decreased spontaneous lung metastasis in tumor-bearing mice. This approach is readily translatable to clinical settings and may increase IR-induced AEs in cancer patients.