Background: Incomplete radiofrequency ablation (iRFA) for hepatocellular carcinoma (HCC) during radiofrequency ablation (RFA) may result in rapid progression of residual tumors and resistance to anti-PD-1 therapy. Research has demonstrated elevated T-cell immunoglobulin and mucin domain 3 (Tim-3) expression in CD8⁺ T cells in the peripheral blood of patients with HCC after RFA, leading to a diminished anti-tumor immune response. Therefore, this study examined the effectiveness of anti-Tim-3 therapy in treating residual tumors after iRFA and explored the underlying mechanisms.

Methods: To examine the expression of Tim-3 in the residual tumors after iRFA for HCC in mice. Treat residual tumors with anti-αTim-3 and evaluate its efficacy. Transcriptomic sequencing was conducted on the residual tumors to explore the underlying mechanisms. Meanwhile, residual tumors were treated with a combination of anti-αTim-3 and anti-αPD-1 to evaluate efficacy.

Results: This study demonstrated elevated Tim-3 expression in CD8⁺ T cells within residual tumors after iRFA. CD8⁺ T cells exhibit attenuated anti-tumor immune responses associated with accelerated tumor progression. Treatment with anti-αTim-3 impeded the advancement of residual tumors by enhancing CD8⁺ T cell infiltration and stimulating their anti-tumor activities. Furthermore, anti-αTim-3 therapy upregulated PD-1 expression in residual tumors. Combination therapy involving anti-αTim-3 and anti-αPD-1 elicited a robust anti-tumor immune response.

Conclusions: Tim-3 expression is elevated in CD8⁺ T cells within residual tumors after iRFA, which contributed to their accelerated advancement. Anti-αTim-3 slows tumor progression by boosting CD8⁺ T cell anti-tumor activity and enhancing response to anti-αPD-1 treatment.

Keywords: Incomplete radiofrequency ablation; PD-1; Tim-3; hepatocellular carcinoma; immunotherapy.

背景: 在射频消融（RFA）治疗肝细胞癌（HCC）时，不完全的射频消融（iRFA）可能导致残留肿瘤迅速进展和对抗PD-1疗法产生抵抗。研究表明，在接受RFA后的HCC患者外周血中，CD8⁺ T细胞中的T细胞免疫球蛋白和粘液素结构域3 (Tim-3)表达升高，导致抗肿瘤免疫反应减弱。因此，本研究考察了抗Tim-3疗法在iRFA后治疗残留肿瘤的有效性，并探讨其潜在机制。

方法: 为了检查小鼠在接受HCC的iRFA后的残留肿瘤中Tim-3的表达情况。用抗αTim-3处理残留肿瘤并评估其疗效。进行转录组测序以探索潜在机制。同时，联合使用抗αTim-3和抗αPD-1治疗残留肿瘤，并评估其效果。

结果: 本研究显示，在iRFA后的HCC残留肿瘤中CD8⁺ T细胞的Tim-3表达升高。CD8⁺ T细胞表现出与肿瘤加速进展相关的抗肿瘤免疫反应减弱。用抗αTim-3治疗可通过增强CD8⁺ T细胞浸润并刺激其抗肿瘤活性来阻碍残留肿瘤的发展。此外，抗αTim-3疗法在残留肿瘤中上调了PD-1表达。联合使用抗αTim-3和抗αPD-1的治疗方案产生了强大的抗肿瘤免疫反应。

结论: iRFA后的HCC残留肿瘤中的CD8⁺ T细胞中Tim-3表达升高，导致其加速发展。抗αTim-3通过增强CD8⁺ T细胞的抗肿瘤活性和提高对抗αPD-1治疗的反应来减缓肿瘤进展。

关键词: 不完全射频消融；PD-1；Tim-3；肝细胞癌；免疫疗法。