Hypothermic oxygenated and normothermic machine perfusion mitigate innate systemic and hepatic inflammation after liver transplantation

Gut, pp. gutjnl–2026

Abstract

BACKGROUND: Immune-mediated injury drives ischaemia-reperfusion injury (IRI) following liver transplantation. While hypothermic oxygenated (HOPE) and normothermic machine perfusion (NMP) improve clinical outcomes, their immunological effects in humans remain unclear. OBJECTIVE: To examine hepatic and systemic immune responses across preservation strategies in a randomised controlled clinical setting. DESIGN: In this mechanistic substudy of the HOPE-NMP randomised controlled trial (NCT04644744), we analysed immune responses in n=47 recipients of extended criteria donor livers from brain-dead donors randomised to static cold storage (SCS), end-ischaemic HOPE or end-ischaemic NMP. Liver biopsies were obtained at organ arrival and early IRI. Blood samples were collected preoperatively, during reperfusion and on postoperative days 1, 2, 3 and 7. METHODS: Analyses included spectral flow cytometry, multiplex immunofluorescence with spatial analyses, cytokine profiling, cellular bioenergetics and mass spectrometry. RESULTS: In human liver allografts, both HOPE and NMP reduced neutrophil infiltration during early IRI, without altering intrahepatic spatial distribution. Machine perfusion induced phenotypic modulation of graft-infiltrating myeloid cells, characterised by downregulation of pro-inflammatory and adhesion markers. Peripheral lymphocyte numbers, particularly of NK cells and T cells, normalised faster after HOPE, compared with SCS and NMP groups, correlating with reduced liver injury. Absolute and relative systemic neutrophil numbers were lower in HOPE-recipients. Machine perfusion decreased the inflammatory activation of circulating myeloid and innate lymphoid cells, while inhibitory immune checkpoints were upregulated postoperatively. CONCLUSIONS: In a randomised controlled trial using human samples, HOPE and NMP differentially modulate hepatic and systemic immune responses, attenuating neutrophil-driven inflammation and promoting immune regulation after liver transplantation.

Authors 27

1. Isabella Lurje

   Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

2. Paul M. Horn

   Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   BIH Biomedical Innovation Academy, BIH Charité Digital Clinician Scientist Program, Berlin Institute of Health, Berlin, Germany

   Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

3. Adrien Guillot

   Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

4. Deniz Uluk

   Heidelberg University · University Hospital Heidelberg

   Affiliation as printed

   Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, Heidelberg, Germany

5. Frederik Schliephake

   Heidelberg University · University Hospital Heidelberg

   Affiliation as printed

   Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, Heidelberg, Germany

6. Luna Rohm

   Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany

7. Dina Katharina Biegel

   Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany

8. Malte Lehnert

   Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany

9. Katharina Remih Aachen Department of Internal Medicine III

   RWTH Aachen University

   Affiliation as printed

   Department of Internal Medicine III, RWTH Aachen, Aachen, Germany

10. Rabea Hokamp Aachen Department of Internal Medicine III

    RWTH Aachen University

    Affiliation as printed

    Department of Internal Medicine III, RWTH Aachen, Aachen, Germany

11. Wiebke Werner

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

12. Alix Bruneau

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

13. Justus Pein

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

    Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany

14. Yaroslava Shevchenko

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

15. Ingrid Wei Zhang

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

16. Pavitra Kumar

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

17. Cornelius Engelmann

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

18. Moritz Peiseler

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

19. Janina Eden

    Medizinische Hochschule Hannover

    Affiliation as printed

    Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany

20. Philipp Dutkowski

    University Hospital Zurich

    Affiliation as printed

    Department of Surgery and Transplantation, Swiss HPB Centre, University Hospital Zurich, Zürich, Switzerland

21. David Meierhofer

    Max Planck Institute for Molecular Genetics

    Affiliation as printed

    Mass Spectrometry Facility, Max-Planck-Institute for Molecular Genetics, Berlin, Germany

22. Nadine Therese Gaisa

    Universität Ulm · University Hospital Ulm

    Affiliation as printed

    Department of Pathology, Ulm University Hospital, Ulm, Germany

23. Pavel Strnad Aachen Department of Internal Medicine III

    Medizinische Universität Lausitz - Carl Thiem · RWTH Aachen University

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Medical University of Lausitz - Carl Thiem, Cottbus, Germany

    Department of Internal Medicine III, RWTH Aachen, Aachen, Germany

24. Johann Pratschke

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany

25. Linda Hammerich

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

26. Frank Tacke

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

27. Georg Lurje corresponding

    Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Gastroenterology and Hepatology, Campus Charité Mitte, Campus Virchow Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany

    Department of Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany