Ajith Ananya, Evraerts Jonathan, Bouzin Caroline, Brusa Davide, Merimi Makram, Najar Mehdi, Smets Françoise, Sokal Etienne, Najimi Mustapha
Laboratory of Pediatric Hepatology and Cell Therapy, Institute of Experimental and Clinical Research, Université catholique de Louvain (UCLouvain), Brussels, Belgium.
Institut de Recherche Expérimentale et Clinique (IREC) Imaging Platform (2IP), Institute of Experimental and Clinical Research, Université catholique de Louvain (UCLouvain), Brussels, Belgium.
Front Immunol. 2025 Jul 8;16:1601215. doi: 10.3389/fimmu.2025.1601215. eCollection 2025.
Hepatocellular carcinoma (HCC) is a classic inflammation related cancer with most cases arising from chronic liver disease (CLD). This study investigates immune dysregulation that occurs during the progression of CLD to HCC by delineating changes in immune cell composition and distribution within the liver microenvironment.
Mice were injected with Diethylnitrosamine (DEN) at 4 weeks of age, followed by continuous tri-weekly injections of carbon tetrachloride (CCl) for 6 and 21 weeks to induce liver fibrosis and HCC. Naïve and Phosphate-buffered saline (PBS) corn oil treated mice were used as controls. Immune cell profiling was performed using multiplex immunofluorescence and flow cytometry analyses.
The spatial analysis of immune cell populations in HCC reveals stable leukocytes overall, with notable increases in myeloid cells, particularly infiltrating macrophages (Inf mph). Indeed, Inf mph show a progressive enrichment from control to tumor, reaching a 5-fold and 10-fold increase in the invasive margin (IM) and surrounding non-tumor tissue (NTT) regions, respectively. T lymphocytes, especially CD4+ T cells but not CD8+ T cells, significantly expand, with CD4+ cells increasing up to 10-fold in the IM and NTT regions of HCC livers. Regulatory T cells (Tregs) population exhibits an extraordinary 125-fold and 80-fold surge in the IM and NTT regions, respectively.
The DEN-CCl induced HCC mouse model replicates key immunosuppressive features of human HCC, notably increased Tregs and macrophages, which provides a robust platform for testing immunotherapies. The prominence of immune cells in the IM region underscores its importance as a critical interface modulating tumor-immune interactions, while the elevated immune presence in the NTT region reflects broader immune dysregulation associated with advanced CLD, and potentially facilitating tumor progression.
肝细胞癌（HCC）是一种典型的炎症相关癌症，大多数病例源于慢性肝病（CLD）。本研究通过描绘肝微环境中免疫细胞组成和分布的变化，调查CLD进展为HCC过程中发生的免疫失调。
4周龄小鼠注射二乙基亚硝胺（DEN），随后连续三周一次注射四氯化碳（CCl），持续6周和21周以诱导肝纤维化和HCC。未处理和用磷酸盐缓冲盐水（PBS）玉米油处理的小鼠用作对照。使用多重免疫荧光和流式细胞术分析进行免疫细胞谱分析。
HCC中免疫细胞群体的空间分析显示总体白细胞稳定，髓系细胞显著增加，特别是浸润性巨噬细胞（Inf mph）。事实上，Inf mph从对照到肿瘤呈现逐渐富集，在侵袭边缘（IM）和周围非肿瘤组织（NTT）区域分别增加了5倍和10倍。T淋巴细胞，尤其是CD4 + T细胞而非CD8 + T细胞显著扩增，CD4 +细胞在HCC肝脏的IM和NTT区域增加高达10倍。调节性T细胞（Tregs）群体在IM和NTT区域分别出现了高达125倍和80倍的激增。
DEN - CCl诱导的HCC小鼠模型复制了人类HCC的关键免疫抑制特征，特别是Tregs和巨噬细胞增加，这为测试免疫疗法提供了一个强大的平台。IM区域免疫细胞的突出强调了其作为调节肿瘤 - 免疫相互作用的关键界面的重要性，而NTT区域免疫存在的升高反映了与晚期CLD相关的更广泛的免疫失调，并可能促进肿瘤进展。
