Spatial organization of neutrophil and macrophage populations in surgically treated hepatocellular carcinoma: a practical tissue-based immunophenotyping study
8 affiliations
- 1Division of Gastroenterology and Hepatology, First Department of Internal Medicine, AHEPA University Hospital, Aristotle University of Thessaloniki
- 2Department of Pathology, School of Medicine, Aristotle University of Thessaloniki
- 3Laboratory of Anatomy, Medical School, Democritus University of Thrace
- 4Department of Physiology, Medical School, National and Kapodistrian University of Athens
- 5Department of Transplantation Surgery, Hippokration General Hospital, School of Medicine, Aristotle University of Thessaloniki
- 6Laboratory of Molecular Hematology, Democritus University of Thrace
- 7Department of Pathology, Democritus University of Thrace
- 8Department of Pathology, Hippokration General Hospital, School of Medicine, Aristotle University of Thessaloniki
Abstract
Background and aims: Hepatocellular carcinoma (HCC) has a spatially heterogeneous immune microenvironment, but the compartment-specific distribution of tumor-associated neutrophils (TANs) and its relationship with systemic inflammation remain incompletely defined. This study characterized CD177-defined neutrophil populations and other immune components across HCC tissue compartments and examined their associations with peripheral neutrophil-to-lymphocyte ratio (NLR).
Methods: Paired tumor center (TC), tumor margin/interface (IM), and adjacent non-neoplastic liver (NNL) specimens from 28 patients with HCC were analyzed using immunofluorescence microscopy and quantitative immunohistochemistry. Total neutrophil elastase-positive (NE ⁺ ) and CD177 ⁺ NE ⁺ neutrophils, CD8 ⁺ lymphocytes, CD68 ⁺ and CD163 ⁺ macrophages, and CD34 ⁺ microvascular density were quantified.
Results: Total NE ⁺ neutrophil density differed across compartments and was higher in both tumor-associated compartments than in NNL (median: TC, 46.40 cells/mm²; IM, 39.65 cells/mm²; NNL, 8.80 cells/mm²; P < 0.001). CD177 ⁺ NE ⁺ density showed a modest compartmental effect and was higher in the TC than at the IM (RR 2.68, 95% CI 1.20–6.02; Holm-adjusted P = 0.050). In contrast, the CD177 ⁺ fraction among total NE ⁺ neutrophils was markedly lower in both tumor-associated compartments than in NNL (median: TC, 17.63%; IM, 10.90%; NNL, 88.08%; P < 0.001)among, accompanied by substantial enrichment of estimated NE ⁺ CD177 ⁻ neutrophils. CD8 ⁺ lymphocytes preferentially accumulated at the IM, whereas CD34 ⁺ microvascular density followed a TC>IM>NNL gradient. CD68 ⁺ macrophages were more abundant in NNL, while CD163 ⁺ macrophage density showed a modest overall compartment effect (mixed-effects P = 0.037), and the CD163/CD68 macrophage-polarization index was higher in the TC than at the IM ( P = 0.009). In exploratory analyses, higher preoperative NLR was associated with the CD163/CD68 macrophage-polarization index specifically in the TC (ρ = 0.451, P = 0.016; q = 0.048), but not with neutrophil abundance, CD177-defined distribution, or patient-level neutrophil spatial-distribution indices.
Conclusions: HCC exhibits marked spatial myeloid heterogeneity, characterized by neutrophil enrichment in tumor-associated compartments, compartmental differences in CD177-defined populations, and compartment-specific macrophage polarization. These findings, generated using a practical tissue-based immunophenotyping approach applicable to routinely processed specimens, warrant validation in larger mechanistic studies.
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BibTeX
@article{arvanitakis2026spatial,
title = {{Spatial organization of neutrophil and macrophage populations in surgically treated hepatocellular carcinoma: a practical tissue-based immunophenotyping study}},
author = {Konstantinos Arvanitakis and Christos Topalidis and Vasileios Papadopoulos and Eirini Giannousi and Georgios Kalopitas and Antonios Chatzigeorgiou and Georgios Katsanos and Anastasia Maria Natsi and Ioannis Mitroulis and Stella Arelaki and Chrysoula Gouta and Evangelia Katsiki and Prodromos Hytiroglou and Georgios Tsoulfas and Triantafyllia Koletsa and Georgios Germanidis},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1982507},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1982507/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Spatial organization of neutrophil and macrophage populations in surgically treated hepatocellular carcinoma: a practical tissue-based immunophenotyping study
AU - Konstantinos Arvanitakis
AU - Christos Topalidis
AU - Vasileios Papadopoulos
AU - Eirini Giannousi
AU - Georgios Kalopitas
AU - Antonios Chatzigeorgiou
AU - Georgios Katsanos
AU - Anastasia Maria Natsi
AU - Ioannis Mitroulis
AU - Stella Arelaki
AU - Chrysoula Gouta
AU - Evangelia Katsiki
AU - Prodromos Hytiroglou
AU - Georgios Tsoulfas
AU - Triantafyllia Koletsa
AU - Georgios Germanidis
PY - 2026
DA - 2026/10/07
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1982507
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1982507/full
AB - Background and aims: Hepatocellular carcinoma (HCC) has a spatially heterogeneous immune microenvironment, but the compartment-specific distribution of tumor-associated neutrophils (TANs) and its relationship with systemic inflammation remain incompletely defined. This study characterized CD177-defined neutrophil populations and other immune components across HCC tissue compartments and examined their associations with peripheral neutrophil-to-lymphocyte ratio (NLR). Methods: Paired tumor center (TC), tumor margin/interface (IM), and adjacent non-neoplastic liver (NNL) specimens from 28 patients with HCC were analyzed using immunofluorescence microscopy and quantitative immunohistochemistry. Total neutrophil elastase-positive (NE ⁺ ) and CD177 ⁺ NE ⁺ neutrophils, CD8 ⁺ lymphocytes, CD68 ⁺ and CD163 ⁺ macrophages, and CD34 ⁺ microvascular density were quantified. Results: Total NE ⁺ neutrophil density differed across compartments and was higher in both tumor-associated compartments than in NNL (median: TC, 46.40 cells/mm²; IM, 39.65 cells/mm²; NNL, 8.80 cells/mm²; P < 0.001). CD177 ⁺ NE ⁺ density showed a modest compartmental effect and was higher in the TC than at the IM (RR 2.68, 95% CI 1.20–6.02; Holm-adjusted P = 0.050). In contrast, the CD177 ⁺ fraction among total NE ⁺ neutrophils was markedly lower in both tumor-associated compartments than in NNL (median: TC, 17.63%; IM, 10.90%; NNL, 88.08%; P < 0.001)among, accompanied by substantial enrichment of estimated NE ⁺ CD177 ⁻ neutrophils. CD8 ⁺ lymphocytes preferentially accumulated at the IM, whereas CD34 ⁺ microvascular density followed a TC>IM>NNL gradient. CD68 ⁺ macrophages were more abundant in NNL, while CD163 ⁺ macrophage density showed a modest overall compartment effect (mixed-effects P = 0.037), and the CD163/CD68 macrophage-polarization index was higher in the TC than at the IM ( P = 0.009). In exploratory analyses, higher preoperative NLR was associated with the CD163/CD68 macrophage-polarization index specifically in the TC (ρ = 0.451, P = 0.016; q = 0.048), but not with neutrophil abundance, CD177-defined distribution, or patient-level neutrophil spatial-distribution indices. Conclusions: HCC exhibits marked spatial myeloid heterogeneity, characterized by neutrophil enrichment in tumor-associated compartments, compartmental differences in CD177-defined populations, and compartment-specific macrophage polarization. These findings, generated using a practical tissue-based immunophenotyping approach applicable to routinely processed specimens, warrant validation in larger mechanistic studies.
SN - 1664-3224
ER -