Cell-free DNA levels are increased in acute graft-versus-host disease

Anna Kroeze, Anne S. Cornelissen, M. Fernanda Pascutti, Myrddin Verheij, Ingrid Bulder, Sjoerd Klarenbeek, Aicha Ait Soussan, Mette D. Hazenberg, Erfan Nur, C. Ellen van der Schoot, Carlijn Voermans, Sacha S. Zeerleder

Research output: Contribution to journalArticleAcademicpeer-review


Background: Cell-free DNA (cfDNA) and nucleosomes, consisting of cfDNA and histones, are markers of cell activation and damage. In systemic inflammation these markers predict severity and fatality. However, the role of cfDNA in acute Graft-versus-Host Disease (aGvHD), a major complication of allogeneic hematopoietic stem cell transplantation (HSCT), is unknown. Objective: The aim of this study is to investigate the role of cfDNA as a marker of aGvHD. Methods: We followed nucleosome levels in 37 allogeneic HSCT patients and an established xenotransplantation mouse model. We determined the origin of cfDNA with a species-specific polymerase chain reaction. Results: In the plasma of aGvHD patients, nucleosome levels significantly increased around the time of aGvHD diagnosis compared to pretransplant, concurrently with a significant increase of known aGvHD markers ST2 and REG3α. In mice, we confirmed that nucleosomes were elevated during clinically detectable aGvHD. We found cfDNA to be mainly of human origin and to a lesser extent of mouse origin, indicating that cfDNA is released by (proliferating) human xeno-reactive PBMC and damaged mouse cells. Conclusion: We show increased cfDNA both in an aGvHD mouse model and in aGvHD patients. We also demonstrate that donor hematopoietic cells and to a lesser degree (damaged) host cells are the cellular source of cfDNA in aGvHD. We propose that nucleosomes and cfDNA might be an additive marker for aGvHD.
Original languageEnglish
Pages (from-to)271-281
Number of pages11
JournalEuropean journal of haematology
Issue number3
Early online date2022
Publication statusPublished - 1 Sep 2022


  • acute graft versus host disease
  • allogeneic hematopoietic stem cell transplantation
  • biomarker
  • cell-free nucleic acids
  • nucleosomes

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