School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Neurosurgery Department, Ruijin Hospital,School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Int J Hyperthermia. 2020;37(1):843-853. doi: 10.1080/02656736.2020.1788173.
In our previous study, a novel cryo-thermal therapy that could stimulate the maturation of innate immune cells to subsequently activate the CD4Th1 cell-dominated antitumor response was developed. However, why cryo-thermal therapy can induce the maturation of innate immunity remains unknown.
In this study, western blot and ELISA were used to analyze the levels of damage-associated molecular patterns (DAMPs, including heat shock protein 70 (HSP70), calreticulin and high-mobility group box protein 1) and in the peripheral blood at different times after cryo-thermal therapy or traditional radiofrequency ablation. The effects of these three DAMPs on myeloid-derived suppressor cells (MDSCs), dendritic cells (DCs) and macrophages were investigated by antibody neutralization . The phenotypic and functional changes in MDSCs, DCs and macrophages were analyzed using FACS and qRT-PCR. An anti-HSP70 antibody was injected intravenously at 6 h after cryo-thermal therapy on days 1 and 2 and mouse survival was monitored.
Cryo-thermal therapy could trigger the release of DAMPs and in the peripheral circulation, which could downregulate the proportion and suppressive signature of MDSCs, and promote the M1 macrophages polarization and DCs maturation. Among three DAMPs, HSP70 played the most evident role in M1 macrophage polarization. neutralization of HSP70 in the early stage of treatment could significantly decrease the survival rate of cryo-thermal therapy treated mice.
Local cryo-thermal therapy not only destroyed solid tumors thermally and mechanically but also induced the release of a large amount of DAMPs to effectively trigger a systemic antitumor response.
在我们之前的研究中，开发了一种新型的冷冻-热疗，它可以刺激固有免疫细胞的成熟，进而激活 CD4Th1 细胞主导的抗肿瘤反应。然而，冷冻-热疗为何能诱导固有免疫的成熟尚不清楚。
本研究采用 Western blot 和 ELISA 分析冷冻-热疗或传统射频消融后不同时间外周血中损伤相关分子模式（DAMPs，包括热休克蛋白 70（HSP70）、钙网蛋白和高迁移率族蛋白 1）的水平。通过抗体中和研究这三种 DAMPs 对髓系来源抑制细胞（MDSCs）、树突状细胞（DCs）和巨噬细胞的影响。采用 FACS 和 qRT-PCR 分析 MDSCs、DCs 和巨噬细胞的表型和功能变化。在冷冻-热疗后 6 小时，第 1 天和第 2 天经静脉注射抗 HSP70 抗体，监测小鼠的存活情况。
冷冻-热疗可触发 DAMPs 的释放，并在外周循环中释放，从而下调 MDSCs 的比例和抑制特征，并促进 M1 巨噬细胞极化和 DCs 的成熟。在这三种 DAMPs 中，HSP70 在 M1 巨噬细胞极化中起最明显的作用。在治疗早期中和 HSP70 可显著降低冷冻-热疗治疗小鼠的存活率。
局部冷冻-热疗不仅能通过热和机械作用破坏实体瘤，还能诱导大量 DAMPs 的释放，从而有效引发全身性抗肿瘤反应。
