Objective: Dendritic cells (DCs) are the most potent antigen-presenting cells, serving as a bridge between innate and adaptive immunity. Activation of the stimulator of interferon genes (STING) pathway by pathogen-derived DNA induces type I interferon responses and promotes CD8+cytotoxic T cell activity. This study aimed to establish a protocol for generating immature DCs from murine bone marrow, optimize their maturation in vitro with a STING agonist, and evaluate their ability to prime naïve T cells for potential use in cancer immunotherapy. Methods: Bone marrow cells from C57BL/6 mice were differentiated into immature DCs under growth factor–supplemented conditions. Maturation was induced using a STING agonist and B16 tumor-derived DNA. Naïve CD4+and CD8+T cells were isolated via magnetic-activated cell sorting (MACS) and co-cultured with the stimulated DCs. Culture conditions were optimized to enhance DC maturation efficiency, and T cell proliferation was assessed following co-culture. Results: Optimization of the culture system markedly increased the yield of mature DCs. Importantly, co-culture of STING agonist-stimulated DCs with naïve T cells resulted in strong CD8+T cell proliferation, indicating effective priming. Conclusions: These findings demonstrate the feasibility of generating functional DCs in vitro and highlight their capacity to prime T cells through STING pathway activation. This proof-of-concept supports the development of DC-based platforms as a promising strategy for novel cancer immunotherapies.
目的：树突状细胞（DCs）是最有效的抗原呈递细胞，在先天免疫与适应性免疫之间起桥梁作用。病原体来源的DNA通过激活干扰素基因刺激因子（STING）通路可诱导I型干扰素反应并增强CD8+细胞毒性T细胞活性。本研究旨在建立从小鼠骨髓中获取未成熟DCs的方案，通过STING激动剂优化其体外成熟过程，并评估其激活初始T细胞的能力，以探索其在癌症免疫治疗中的应用潜力。方法：在生长因子补充条件下，将C57BL/6小鼠骨髓细胞分化为未成熟DCs，采用STING激动剂及B16肿瘤来源DNA诱导其成熟。通过磁激活细胞分选技术（MACS）分离初始CD4+与CD8+T细胞，并与活化的DCs共培养。通过优化培养条件提升DCs成熟效率，并在共培养后评估T细胞增殖情况。结果：培养体系的优化显著提高了成熟DCs的产量。关键发现显示，经STING激动剂刺激的DCs与初始T细胞共培养后，能有效诱导CD8+T细胞显著增殖，表明其具备高效的抗原呈递能力。结论：本研究证实了体外制备功能性DCs的可行性，并揭示了通过激活STING通路DCs能够有效激活T细胞。这一概念验证为开发基于DCs的新型癌症免疫治疗平台提供了理论依据。
In Vitro Maturation of Bone Marrow-Derived Dendritic Cells via STING Activation for T Cell Priming
肿瘤(癌症)患者之家