Cryoablation remodels the immune microenvironment in hepatocellular carcinoma: From mechanistic insights to clinical translation in combination immunotherapy

Table 4 Summary of key preclinical studies on cryoablation combined with immunomodulatory strategies

Ref.	Disease model	Ablation completeness	Combination regimen and sequencing	Sample size	Key immunological and therapeutic outcomes
Mandt et al[27], 2023	Murine HCC	Incomplete	CpG (Toll-like receptor 9 agonist) + αPD-1, administered after cryoablation	n = 63 total	Synergistically enhanced systemic antitumor immunity, characterized by increased intratumoral CD8+ T cell infiltration and a robust Th1-type cytokine response, leading to inhibited distal tumor growth and prolonged survival
Gu et al[28], 2024	Murine HCC	Complete	Anti-PD-1 + anti-CTLA-4 (dual immune checkpoint inhibitor), concurrent with/after cryoablation	n = 40 total, n = 10/group	Most effectively remodeled the TME by promoting CD8+ and CD4+ T cell infiltration while reducing regulatory T cells and myeloid-derived suppressor cells, resulting in potent inhibition of distant tumors and a synergistic effect
Qian et al[29], 2024	Bilateral subcutaneous HCC	Complete	αPD-1 + αCTLA-4, after cryo-thermal ablation	Not specified	Converted immunologically “cold” tumors to “hot” by inducing immunogenic cell death and enhancing DC activation, with the triple combination eliciting the strongest abscopal effect
Shewarega et al[37], 2024	Murine HCC	Incomplete	Matrix metalloproteinase inhibitor, administered after cryoablation	n = 40 total, n = 6/group	Specifically increased intratumoral CD8+ T cell infiltration, providing a model to study the immune effects of subtotal ablation
Ghani et al[38], 2023	Murine HCC	Incomplete	CPMV priming, followed by cryoablation, and then a CPMV boost	n = 44-56 total, n = 11-14/group	Functioned as an in situ vaccine, where CPMV acted as a potent innate immune stimulant, synergizing with cryoablation-released antigens to enhance antigen-presenting cell recruitment and cross-priming
Yang et al[89], 2025	Murine cervical cancer	Complete	αPD-1, administered after cryoablation	Bilateral model; typical n = 3-4/group for assays	Induced an effective abscopal effect by remodeling the TME, increasing cytotoxic lymphocyte activity, and upregulating programmed death-ligand 1 expression, thereby sensitizing tumors to checkpoint blockade
Wang and Guo[90], 2025	Murine colorectal cancer liver metastasis	Complete (described as tumor fully ice-covered)	Cryoablation + granulocyte-macrophage colony-stimulating factor (administered immediately post-ablation)	n = 80 total, n = 20/group	Enhanced antitumor immune response: Significantly and persistently increased intratumoral infiltration of CD11c+ DCs; promoted a Th1-biased immune response with sustained high serum interferon-gamma levels and suppressed interleukin-4 levels; associated with prolonged survival in the combination group

CPMV: Cowpea mosaic virus; CTL: Cytotoxic T lymphocyte; DC: Dendritic cell; HCC: Hepatocellular carcinoma; PD-1: Programmed death-1; TME: Tumor microenvironment.