Abstract

Purpose

In our previous study, a novel cryo-thermal therapy that could stimulate the maturation of innate immune cells to subsequently activate the CD4⁺Th1 cell-dominated antitumor response was developed. However, why cryo-thermal therapy can induce the maturation of innate immunity remains unknown.

Methods

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) in situ 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 in vitro. 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.

Results

Cryo-thermal therapy could trigger the release of DAMPs in situ 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. In vivo neutralization of HSP70 in the early stage of treatment could significantly decrease the survival rate of cryo-thermal therapy treated mice.

Conclusions

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.

中文翻译：

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低温热疗法后释放的肿瘤相关 HSP70 靶向抗肿瘤免疫反应中的先天免疫启动。

摘要

目的

在我们之前的研究中，开发了一种新的低温热疗法，可以刺激先天免疫细胞的成熟，随后激活 CD4 ⁺ Th1 细胞主导的抗肿瘤反应。然而，为什么低温热疗法可以诱导先天免疫的成熟仍然未知。

方法

在本研究中，蛋白质印迹和 ELISA 用于分析原位和外周血中损伤相关分子模式（DAMP，包括热休克蛋白 70 (HSP70)、钙网蛋白和高迁移率组框蛋白 1）的水平。冷热疗法或传统射频消融后的不同时间。通过体外抗体中和研究了这三种 DAMP 对髓源性抑制细胞 (MDSC)、树突状细胞 (DC) 和巨噬细胞的影响。使用 FACS 和 qRT-PCR 分析 MDSC、DC 和巨噬细胞的表型和功能变化。在第 1 天和第 2 天低温热疗法后 6 小时静脉内注射抗 HSP70 抗体，并监测小鼠存活率。

结果

低温热疗法可触发原位和外周循环中 DAMP 的释放，从而下调 MDSCs 的比例和抑制信号，促进 M1 巨噬细胞极化和 DCs 成熟。在三种 DAMP 中，HSP70 在 M1 巨噬细胞极化中的作用最为明显。HSP70在治疗早期的体内中和可显着降低冷冻热疗法治疗小鼠的存活率。

结论

局部低温热疗法不仅可以热力和机械地破坏实体瘤，还可以诱导大量 DAMP 的释放，从而有效地引发全身性抗肿瘤反应。