ABSTRACT

Microwave ablation (MWA) has been widely used in the treatment of solid tumors. Studies have been less conducted on the efficacy of MWA used with cell immunotherapy in treating hepatocellular carcinoma (HCC). The current study aimed at exploring the efficacy of MWA in combination with cell immunotherapy in treating HCC. Hepa1-6 HCC mice were treated by MWA, blockade, or the combined therapy (MWA used with blockade), or left untreated. Survival rates of the mice were plotted by Kaplan-Meier Curve, followed by log-rank test. 25 days after the operation, surviving mice were monitored for tumor recurrence, and tumor volumes were calculated every 5 days. Immunohistochemistry and flow cytometry were performed to detect the numbers of CD4⁺ and CD8⁺ cells in the tumors and spleens of mice. The expressions of related cytokines were detected and measured by ELISPOT and ELISA. The results showed that MWA combined with anti-PD-1/anti-CTLA-4 not only increased the survival time, protected the mice against tumor recurrence, but also enhanced the intratumoral infiltration of cytotoxic T lymphocyte and systemic T-cell immune responses induced by MWA through activation of synergistically specific antitumor immunity. In addition, the combined therapy increased T-helper 1 cell (Th1-type) cytokines, but reduced Th2-type cytokines, resulting in the polarization of Th1 cells. T-cell immune responses of HCC cells were activated by MWA. In addition, the combined therapy of MWA and anti-PD-1/anti-CTLA-4 induced Th1-type immune response, and showed specific antitumor immunity.

摘要

微波消融（MWA）已广泛应用于实体瘤的治疗。关于 MWA 与细胞免疫疗法一起用于治疗肝细胞癌 (HCC) 的功效的研究较少。目前的研究旨在探索 MWA 联合细胞免疫疗法治疗 HCC 的疗效。Hepa1-6 HCC 小鼠通过 MWA、阻断或联合疗法（MWA 与阻断一起使用）进行治疗，或不进行治疗。通过 Kaplan-Meier 曲线绘制小鼠的存活率，然后进行对数秩检验。术后25天，监测存活小鼠的肿瘤复发情况，每5天计算一次肿瘤体积。进行免疫组化和流式细胞术检测CD4 ⁺和CD8 ^{+ 的数量}小鼠肿瘤和脾脏中的细胞。通过ELISPOT和ELISA检测和测量相关细胞因子的表达。结果表明，MWA联合anti-PD-1/anti-CTLA-4不仅增加了存活时间，保护了小鼠免于肿瘤复发，而且增强了细胞毒性T淋巴细胞的肿瘤内浸润和全身T细胞免疫反应。 MWA 通过激活协同特异性抗肿瘤免疫。此外，联合治疗增加了T辅助1细胞（Th1型）细胞因子，但减少了Th2型细胞因子，导致Th1细胞极化。MWA 激活 HCC 细胞的 T 细胞免疫反应。此外，MWA与抗PD-1/抗CTLA-4联合治疗可诱导Th1型免疫反应，表现出特异性抗肿瘤免疫。