【摘要】 目的：探讨基因工程化细菌囊泡搭载光敏剂Ce6的抗肿瘤效果。方法：通过基因工程手段使大肠埃希菌Escherichia coli Nissle 1917(EcN)过表达干扰素α(IFNα)并提取相应细菌囊泡（BVs-IFNα），通过超声将光敏剂Ce6装载在细菌囊泡表面得到BVs-IFNα@Ce6纳米颗粒。运用动态光散射、纳米颗粒跟踪技术共聚焦成像及紫外吸收光谱对BVs-IFNα@Ce6复合体进行表征。将得到的BVs-IFNα@Ce6纳米颗粒与小鼠黑色素瘤细胞（B16F10）和小鼠骨髓来源树突状细胞（BMDC）共培养，用CCK-8法评估其光动力治疗效果，用流式细胞术验证其免疫激活能力。结果：成功制备出BVs-IFNα@Ce6纳米颗粒，其粒径在180 nm左右具有稳定性。在红光照射条件下，BVs-IFNα@Ce6对B16F10肿瘤细胞表现出明显的杀伤能力，并且可以有效地促进BMDC细胞表面免疫相关分子MHC-II、CD80/CD86的表达，展现出显著的免疫激活能力。结论：BVs-IFNα@Ce6纳米颗粒兼具增强光动力疗效与激活抗肿瘤免疫的双重功能，具备临床转化潜力。更多还原

【Abstract】 Objective To investigate the antitumor effect of genetically engineered bacterial vesicles loaded with the photosensitizer Ce6. Methods Genetic engineering techniques were used to overexpress interferon α(IFNα) in Escherichia coli Nissle 1917(EcN), and the corresponding bacterial vesicles(BVs-IFNα) were extracted. The photosensitizer Ce6 was loaded onto the surface of the bacterial vesicles via ultrasonication to obtain BVs-IFNα@Ce6 nanoparticles. The BVsIFNα@Ce6 complex was characterized using dynamic light scattering, nanoparticle tracking technology, confocal imaging,and ultraviolet absorption spectroscopy. The obtained BVs-IFNα@Ce6 nanoparticles were co-cultured with mouse melanoma cells(B16F10) and mouse bone marrow-derived dendritic cells(BMDC). The efficacy of photodynamic therapy was assessed using the CCK-8 method, and its ability to induce immune activation was verified using flow cytometry. Results The BVs-IFNα@Ce6 nanoparticles were successfully prepared with a stable particle size of around 180 nm. Under red light irradiation, BVs-IFNα@Ce6 exhibited significant cytotoxicity against B16F10 tumor cells and effectively promoted the expressions of immune-related molecules MHC-II, CD80/CD86 on the surface of BMDC cells, demonstrating its remarkable ability to induce immune activation. Conclusion The BVs-IFNα@Ce6 nanoparticles possess dual functions of enhancing photodynamic therapy efficacy and activating antitumor immunity, showing potential for clinical translation.更多还原