a School of Biomedical Engineering and Med-X Research Institute , Shanghai Jiao Tong University , Shanghai , PR China.
Int J Hyperthermia. 2019;36(1):408-420. doi: 10.1080/02656736.2019.1579373. Epub 2019 Mar 20.
We previously developed a novel cryo-thermal therapy to treat malignant mammary carcinoma and melanoma in a mouse model; long-term survival and CD4 T cell orchestrating anti-tumor immune memory response were achieved. Moreover, cryo-thermal-induced CD4 T cell differentiation into Th1 and CD4CTL sub-lineages, in which M1 macrophage polarization played a direct, important role. In particular, cryo-thermal therapy triggered M1 macrophage polarization with up-regulated expression of C-X-C motif ligand 10 (CXCL10) and Interleukin 6 (IL-6). But whether CXCL10 and IL-6 contribute to CD4 T cell-mediated anti-tumor immunity remains unclear. In this study, the role of cryo-thermal-induced CXCL10 and IL-6 in anti-tumor immunity was determined.
The level of CXCL10 and IL-6 in spleen and serum was determined by RT-PCR and ELISA on day 14 after cryo-thermal therapy. Splenic dendritic cells (DCs) and macrophages were isolated from cryo-thermal-treated mice on day 5 and 14, and the level of CXCL10 and IL-6 in macrophages and DCs was determined by ELISA. The transwell migration assay was performed to study immune cell migration. In vivo neutralization of CXCL10 or IL-6 was performed to investigate the phenotypic changes of immune cells.
Cryo-thermal therapy induced M1 macrophage polarization with up-regulation of CXCL10 and IL-6 expression in spleen. CXCL10 and IL-6 promoted DCs migration and maturation, and subsequently promoted CD4 T cell migration and differentiation into Th1 and CD4 CTL, moreover, reduced myeloid-derived suppressor cells (MDSCs) accumulation.
Cryo-thermal-induced CXCL10 and IL-6 created acute inflammatory environment to initiate a systemically cascading innate and adaptive anti-tumor immunity, which was more permissive for tumor eradication.
我们之前开发了一种新的冷冻-热疗方法来治疗小鼠模型中的恶性乳腺肿瘤和黑色素瘤；实现了长期生存和 CD4 T 细胞协调抗肿瘤免疫记忆反应。此外，冷冻-热诱导的 CD4 T 细胞分化为 Th1 和 CD4CTL 亚群，其中 M1 巨噬细胞极化起着直接的、重要的作用。特别是，冷冻-热疗触发 M1 巨噬细胞极化，上调 C-X-C 基序配体 10（CXCL10）和白细胞介素 6（IL-6）的表达。但是，CXCL10 和 IL-6 是否有助于 CD4 T 细胞介导的抗肿瘤免疫尚不清楚。在这项研究中，确定了冷冻-热诱导的 CXCL10 和 IL-6 在抗肿瘤免疫中的作用。
在冷冻-热治疗后第 14 天，通过 RT-PCR 和 ELISA 测定脾和血清中的 CXCL10 和 IL-6 水平。从冷冻-热治疗的小鼠中分离出第 5 天和第 14 天的脾树突状细胞（DC）和巨噬细胞，并通过 ELISA 测定巨噬细胞和 DC 中 CXCL10 和 IL-6 的水平。进行 Transwell 迁移实验以研究免疫细胞迁移。体内中和 CXCL10 或 IL-6 以研究免疫细胞的表型变化。
冷冻-热治疗诱导 M1 巨噬细胞极化，导致脾中 CXCL10 和 IL-6 表达上调。CXCL10 和 IL-6 促进 DC 迁移和成熟，随后促进 CD4 T 细胞迁移和分化为 Th1 和 CD4CTL，此外，减少骨髓源性抑制细胞（MDSCs）的积累。
冷冻-热诱导的 CXCL10 和 IL-6 创造了急性炎症环境，引发了全身性级联的先天和适应性抗肿瘤免疫，这更有利于肿瘤的清除。
