肿瘤疫苗是癌症免疫治疗的一个有前途的途径。尽管针对特定免疫表位的研究取得了进展，但缺乏这些表位的肿瘤细胞仍可能逃避治疗。在此，我们的目的是构建一种高效的原位肿瘤疫苗Vac-SM，利用紫草素（SKN）诱导免疫原性细胞死亡（ICD）和耻垢分枝杆菌（M. smegmatis）作为免疫佐剂来增强原位肿瘤疫苗的功效。通过CCK-8检测，SKN对肿瘤细胞系表现出剂量依赖性和时间依赖性的细胞毒作用，并通过检测相关指标的表达分别诱导肿瘤细胞中的ICD。与对照组相比，小鼠皮下转移性肿瘤原位注射Vac-SM显着抑制肿瘤生长和远处肿瘤生长，提高生存率。根据流量，耻垢分枝杆菌有效诱导骨髓源性树突状细胞 (DC) 成熟和激活，体内肿瘤引流淋巴结显示，Vac-SM 治疗后 DC 成熟度增加，效应记忆 T 细胞亚群比例更高总的来说，Vac-SM疫苗有效诱导ICD，改善DC的抗原呈递，激活特异性全身抗肿瘤T细胞免疫反应，表现出良好的安全性，并有望用于局部肿瘤免疫治疗的临床转化。

Tumor vaccines are a promising avenue in cancer immunotherapy. Despite the progress in targeting specific immune epitopes, tumor cells lacking them can evade treatment. Here, we aimed to construct an efficient in situ tumor vaccine Vac-SM, utilizing shikonin (SKN) to induce immunogenic cell death (ICD) and Mycobacterium smegmatis ( M. smegmatis) as an immune adjuvant to enhance in situ tumor vaccine efficacy. SKN demonstrated a dose-dependent and time-dependent cytotoxic effect on the tumor cell line as seen using the CCK-8 assay and induced ICD in tumor cells by detecting the expression of relevant indicators respectively. Compared to that in the control groups, in situ Vac-SM injection in mouse subcutaneous metastatic tumors significantly inhibited tumor growth and distant tumor growth and improved survival rates. M. smegmatis effectively induced bone marrow-derived dendritic cells (DC) maturation and activation and in vivo tumor-draining lymph nodes showed increased maturation of DC and a higher proportion of effector memory T-cell subsets with Vac-SM treatment, based on flow cytometry analysis results.Collectively, Vac-SM vaccine effectively induces ICD, improves antigen presentation by DC, activates a specific systemic antitumor T-cell immune response, exhibits favorable safety profile, and holds promise for clinical translation for local tumor immunotherapy.