Sarcosine promotes trafficking of dendritic cells and improves efficacy of anti-tumor dendritic cell vaccines via CXC chemokine family signaling
Sarcosine promotes trafficking of dendritic cells and improves efficacy of anti-tumor dendritic cell vaccines via CXC chemokine family signaling
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DOI：
10.1186/s40425-019-0809-4
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发表时间：
2019-11-21
影响因子：
10.9
通讯作者：
Rahman, Maryam
中科院分区：
文献类型：
--
作者：
Dastmalchi, Farhad;Karachi, Aida;Rahman, Maryam
树突状细胞（DC）疫苗的有效性与DC在运送到患者体内后向淋巴结迁移的效率直接相关。我们发现一种天然存在的代谢物，肌氨酸，增加了DC在人和小鼠细胞中的迁移，从而显著提高了抗肿瘤功效。我们假设肌氨酸诱导的细胞迁移是由于趋化因子信号传导。方法从野生型C57BL/6小鼠骨髓中获取dc，用肿瘤信使RNA （mRNA）电穿孔。从人外周血单个核细胞（PBMCs）中分离出树突状细胞。DCs用20 mM肌氨酸处理。从转基因小鼠中分离抗原特异性T细胞，静脉注射到荷瘤小鼠体内。DC疫苗通过皮内注射提供。用流式细胞术和免疫荧光显微镜观察体内迁移情况。采用RT-PCR和Nanostring技术研究dc中RNA基因的表达。结果肌氨酸能显著促进人和小鼠体外DC迁移。在小鼠体内，经肌氨酸处理的dc在皮内递送后向淋巴结和脾脏的迁移均显著增加。在B16F10-OVA肿瘤侧腹模型中，肌氨酸处理的DC疫苗显著改善了肿瘤控制，并提高了GL261-gp100颅内胶质瘤模型的生存率。在肌氨酸处理的dc中，CXCR2、CXCL3和CXCL1的基因表达上调。进一步的代谢分析表明，环氧化酶-1和Pik3cg表达上调。在人和小鼠dc中加入CXCR2中和抗体可消除肌氨酸诱导的迁移。CXCR2中和抗体也消除了肿瘤模型中肌氨酸处理的dc的生存益处。结论肌氨酸通过CXC趋化因子途径促进小鼠和人dc的迁移。该平台可用于改进现有的DC疫苗策略。
Background Dendritic cell (DC) vaccine efficacy is directly related to the efficiency of DC migration to the lymph node after delivery to the patient. We discovered that a naturally occurring metabolite, sarcosine, increases DC migration in human and murine cells resulting in significantly improved anti-tumor efficacy. We hypothesized that sarcosine induced cell migration was due to chemokine signaling. Methods DCs were harvested from the bone marrow of wild type C57BL/6 mice and electroporated with tumor messenger RNA (mRNA). Human DCs were isolated from peripheral blood mononuclear cells (PBMCs). DCs were treated with 20 mM of sarcosine. Antigen specific T cells were isolated from transgenic mice and injected intravenously into tumor bearing mice. DC vaccines were delivered via intradermal injection. In vivo migration was evaluated by flow cytometry and immunofluorescence microscopy. Gene expression in RNA was investigated in DCs via RT-PCR and Nanostring. Results Sarcosine significantly increased human and murine DC migration in vitro. In vivo sarcosine-treated DCs had significantly increased migration to both the lymph nodes and spleens after intradermal delivery in mice. Sarcosine-treated DC vaccines resulted in significantly improved tumor control in a B16F10-OVA tumor flank model and improved survival in an intracranial GL261-gp100 glioma model. Gene expression demonstrated an upregulation of CXCR2, CXCL3 and CXCL1 in sarcosine- treated DCs. Further metabolic analysis demonstrated the up-regulation of cyclooxygenase-1 and Pik3cg. Sarcosine induced migration was abrogated by adding the CXCR2 neutralizing antibody in both human and murine DCs. CXCR2 neutralizing antibody also removed the survival benefit of sarcosine-treated DCs in the tumor models. Conclusion Sarcosine increases the migration of murine and human DCs via the CXC chemokine pathway. This platform can be utilized to improve existing DC vaccine strategies.