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Original article
Chemokine-driven lymphocyte infiltration: an early intratumoural event determining long-term survival in resectable hepatocellular carcinoma
  1. Valerie Chew1,
  2. Jinmiao Chen1,
  3. Deming Lee1,
  4. Evelyn Loh1,
  5. Joyce Lee2,3,
  6. Kiat Hon Lim4,
  7. Achim Weber5,
  8. Ksenija Slankamenac6,
  9. Ronnie T P Poon3,7,
  10. Henry Yang1,
  11. London Lucien P J Ooi8,9,
  12. Han Chong Toh8,
  13. Mathias Heikenwalder10,11,12,
  14. Irene O L Ng2,3,
  15. Alessandra Nardin1,
  16. Jean-Pierre Abastado1
  1. 1Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore
  2. 2Department of Pathology, The University of Hong Kong, Queen Mary Hospital, Hong Kong, China
  3. 3State Key Laboratory for Liver Research, The University of Hong Kong, Hong Kong, China
  4. 4Department of Pathology, Singapore General Hospital, Singapore, Singapore
  5. 5Department of Clinical Pathology, University Hospital of Zurich, Zurich, Switzerland
  6. 6Department of Visceral and Transplantation Surgery, University Hospital of Zurich, Zurich, Switzerland
  7. 7Department of Surgery, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong
  8. 8Department of Medical Oncology, National Cancer Centre, Singapore, Singapore
  9. 9Department of General Surgery, Singapore General Hospital, Singapore, Singapore
  10. 10Institute of Neuropathology, University Hospital of Zurich, Zurich, Switzerland
  11. 11Institute of Virology, Technical University München, München, Germany
  12. 12Helmholtz Zentrum München, Germany
  1. Correspondence to Jean-Pierre Abastado, Singapore Immunology Network, BMSI, A*STAR, Immunos #04-00, 8A Biomedical Grove, Singapore; abastado{at}


Objective Hepatocellular carcinoma (HCC) is a heterogeneous disease with poor prognosis and limited methods for predicting patient survival. The nature of the immune cells that infiltrate tumours is known to impact clinical outcome. However, the molecular events that regulate this infiltration require further understanding. Here the ability of immune genes expressed in the tumour microenvironment to predict disease progression was investigated.

Methods Using quantitative PCR, the expression of 14 immune genes in resected tumour tissues from 57 Singaporean patients was analysed. The nearest-template prediction method was used to derive and test a prognostic signature from this training cohort. The signature was then validated in an independent cohort of 98 patients from Hong Kong and Zurich. Intratumoural components expressing these critical immune genes were identified by in situ labelling. Regulation of these genes was analysed in vitro using the HCC cell line SNU-182.

Results The identified 14 immune-gene signature predicts patient survival in both the training cohort (p=0.0004 and HR=5.2) and the validation cohort (p=0.0051 and HR=2.5) irrespective of patient ethnicity and disease aetiology. Importantly, it predicts the survival of patients with early disease (stages I and II), for whom classical clinical parameters provide limited information. The lack of predictive power in late disease stages III and IV emphasises that a protective immune microenvironment has to be established early in order to impact disease progression significantly. This signature includes the chemokine genes CXCL10, CCL5 and CCL2, whose expression correlates with markers of T helper 1 (Th1), CD8+ T and natural killer (NK) cells. Inflammatory cytokines (tumour necrosis factor α, interferon γ) and Toll-like receptor 3 ligands stimulate intratumoural production of these chemokines which drive tumour infiltration by T and NK cells, leading to enhanced cancer cell death.

Conclusion A 14 immune-gene signature, which identifies molecular cues driving tumour infiltration by lymphocytes, accurately predicts survival of patients with HCC especially in early disease.

  • Hepatocellular carcinoma
  • immune-gene signature
  • tumour microenvironment
  • chemokines
  • lymphocytes tumour infiltration
  • immunology in hepatology
  • tumour markers
  • liver immunology
  • immunohistochemistry
  • acute hepatitis
  • cancer
  • oncogenes
  • gastric cancer
  • liver metastases
  • pancreatic pathology
  • pancreatic tumours
  • molecular pathology
  • gastrointestinal lymphoma
  • liver biopsy
  • surgical complications
  • surgical oncology
  • surgical resection
  • immune response

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-commercial License, which permits use, distribution, and reproduction in any medium, provided the original work is properly cited, the use is non commercial and is otherwise in compliance with the license. See: and

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  • Funding This study was mainly funded by The Biomedical Sciences Institutes (BMSI), Agency for Science, Technology and Research (A*STAR), and partly funded by a Hong Kong Research Grants Council Collaborative Research Grant to IOLN (HKU 7/CRG/09).

  • Competing interests None.

  • Ethics approval This study was conducted with the approval of the respective Institutional Review Boards.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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