为了测试假设：冷冻消融和豇豆花叶病毒（CPMV）内肿瘤免疫调节纳米颗粒联合作为原位疫苗接近接种在小鼠肝细胞癌（HCC）模型中诱导全身抗肿瘤免疫。小鼠双侧皮下RIL-175细胞源性HCC随机分为4组：PBS（对照组）、仅冷冻消融组（Cryo）、仅CPMV处理组（CPMV）或冷冻消融加CPMV处理组（Cryo+CPMV）（每组N=11-14）。每3天给予肿瘤内CPMV 4次剂量，第3天进行冷冻消融，监测对侧肿瘤。测量肿瘤生长和全身趋化因子/细胞因子浓度。收集部分肿瘤和脾脏进行免疫组化和流式细胞术。进行单向或双向ANOVA进行统计比较，p值<0.05为显著水平。治疗2周后，Cryo和CPMV单独或联合均优于对照组在治疗的肿瘤中，但Cryo+CPMV表现出最强的减少和最低的差异（1.6 ± 0.9倍与6.3 ± 0.5倍；p<0.0001）。对于未治疗的肿瘤，仅Cryo+CPMV明显降低了肿瘤生长，与对照组相比（9.2 ± 0.9倍与17.8 ± 2.1倍；p=0.01）。 Cryo+CPMV组显示未治疗肿瘤中IL-10的短暂增加和持续降低的CXCL1。流式细胞术显示未治疗肿瘤中自然杀伤细胞富集，而脾脏中PD-1表达增加。冷冻消融和肿瘤内CPMV，无论组合还是单独，均显示了对治疗的HCC肿瘤具有强大的疗效，但仅有冷冻消融加CPMV可以减缓未治疗肿瘤的生长，这与绝地效应相符。版权所有©2023。Elsevier Inc.出版。

To test the hypothesis that cryoablation combined with intratumoral immunomodulating nanoparticles from cowpea mosaic virus (CPMV) as an in situ vaccination approach induces systemic antitumoral immunity in a murine model of hepatocellular carcinoma (HCC).Mice with bilateral, subcutaneous RIL-175 cell-derived HCCs were randomized to 4 groups: PBS (Control), Cryoablation only (Cryo), CPMV-treated only (CPMV), or Cryoablation plus CPMV-treated (Cryo+CPMV) (N=11-14/group). Intratumoral CPMV was administered every 3 days for 4 doses, with cryoablation on the third day. Contralateral tumors were monitored. Tumor growth and systemic chemokine/cytokine concentrations were measured. A subset of tumors and spleens were harvested for immunohistochemistry and flow cytometry. One- or two-way ANOVA was performed for statistical comparisons. A p value of <0.05 was the threshold for statistical significance.At 2 weeks post-treatment, Cryo and CPMV, alone or combined, outperformed Control in the treated tumor, but Cryo+CPMV showed the strongest reduction and lowest variance (1.6 ± 0.9-fold vs 6.3 ± 0.5-fold; p<0.0001). For the untreated tumor, only Cryo+CPMV significantly reduced tumor growth vs Control (9.2 ± 0.9-fold vs 17.8 ± 2.1-fold; p=0.01). The Cryo+CPMV group exhibited a transient increase in IL-10 and persistently decreased CXCL1. Flow cytometry revealed natural killer cell enrichment in the untreated tumor and increased PD-1 expression in the spleen. Tumor-infiltrating lymphocytes increased in Cryo+CPMV tumors by IHC.Cryoablation and intratumoral CPMV, together or alone, demonstrated potent efficacy against treated HCC tumors, but only cryoablation combined with CPMV slowed the growth of untreated tumors, consistent with an abscopal effect.Copyright © 2023. Published by Elsevier Inc.