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Peer-reviewedCC BY 4.0Frontiers in Immunology

Composition of immune cells in peripheral blood of patients with leukemia in remission after stem cell transplantation (HSCT) with or without graft-versus-host disease (GvHD): a comparative analysis with healthy donors

  • Vivien M. Simon1,2,3
  • Cansu Karatas1,2
  • Kristina Schreiner1,2
  • Anying Huang1,2
  • Annalena Frischhut1,2
  • Joudi Abdulmajid1,2
  • Christoph Schmid2,4,5
  • Jochen J. Frietsch6
  • Helene Schmeller7
  • Tobias A. W. Holderried7,8
  • Elisa Sala9
  • Helga Schmetzer1,2
2 more authors
  • Giuliano Filippini Velazquez2,4,5
  • Jan Weller10
10 affiliations
  1. 1Department of Medicine III, University Hospital Ludwig-Maximilians-Universität (LMU) Munich
  2. 2Bavarian Cancer Research Center (BZKF)
  3. 3Faculty of Biology, University of Bielefeld
  4. 4Section for Stem Cell Transplantation and Cellular Therapy Research, Faculty of Medicine, University of Augsburg
  5. 5Comprehensive Cancer Center Alliance WERA (CCC WERA)
  6. 6Department of Internal Medicine II, University Hospital of Würzburg
  7. 7University of Bonn, University Hospital Bonn, Medical Clinic and Polyclinic III Internal Medicine with a focus on Oncology, Hematology, and Rheumatology
  8. 8Clinical Division of Hematology, Department of Internal Medicine, Medical University of Graz
  9. 9Department of Hematology and Oncology, University Hospital Ulm
  10. 10Department of Oncology, Hematology and Bone Marrow Transplantation with Division of Pneumology, University Medical Center Hamburg-Eppendorf
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Abstract

Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is potentially curative for many hematologic malignancies. Clinical outcomes are critically determined by the balance between graft-versus-malignancy effects and graft-versus-host disease (GvM and GvHD, respectively). We hypothesized that integrated immune phenotyping can distinguish immune tolerance from clinically relevant alloreactivity in patients with hematological neoplasia who are in remission after HSCT. These findings may have implications for GvHD screening and monitoring and could support treatment decision-making.

Methods: Multi-color flow cytometry of peripheral blood mononuclear cells profiled myeloid, adaptive, and innate compartments in healthy donors ( n = 10) and post-HSCT patients in remission: without GvHD [NoGvHD ( n = 19)], with chronic (c) GvHD ( n = 18, including n = 4 overlap-a/cGvHD), and with acute (a) GvHD ( n = 6). Frequencies of immune cell subsets were compared between cohorts and stratified by clinical features [e.g., severity, organ involvement, donor lymphocyte infusion (DLI), GvHD development, and cGvHD progression].

Results: NoGvHD patients (clinically asymptomatic and whose disease were in remission) showed immune-cell composition differing substantially from healthy donors: CD14 ⁺ monocytes (Mo), M-MDSCs, and especially checkpoint molecule (cpm)-expressing Mo (TIM3, 2B4, and PDL1) were reduced, while CTLA-4 ⁺ polymorphonuclear myeloid-derived suppressor cells (CTLA-4 ⁺ -PMN-MDSCs) were elevated, as well as T-effector memory (Tem) cells and cpm-expressing T cells (PD1, LAG3, 2B4, TIM3, and TIGIT). cGvHD showed eroded regulatory myeloid subsets and skewed T cells toward effector-memory dominance with increased frequencies of inhibitory cpm-expressing T cells. aGvHD showed marked late-proliferating T-cell expansion (T prol-late /T ³⁺ fold change 9.92 vs. Healthy), T reg depletion, and loss of regulatory NK 56bri populations. Overlap a/cGvHD displayed chronic T-cell stimulation with altered tissue homing (Intβ7 ⁺ enriched), and DLI-induced GvHD showed effector differentiation with insufficient regulatory compensation.

Conclusion: NoGvHD is not “Healthy minus disease” but a distinct post-HSCT immune state. Multi-compartment immune signature profiling by flow cytometry distinguishes preserved tolerance (Healthy) from post-HSCT remodeling (NoGvHD), progressive dysregulation (cGvHD), and acute hyperactivation (aGvHD), and refines GvHD subgroups by clinical context.

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Cite

BibTeXRIS
BibTeX
@article{simon2026composition,
  title = {{Composition of immune cells in peripheral blood of patients with leukemia in remission after stem cell transplantation (HSCT) with or without graft-versus-host disease (GvHD): a comparative analysis with healthy donors}},
  author = {Vivien M. Simon and Cansu Karatas and Kristina Schreiner and Anying Huang and Annalena Frischhut and Joudi Abdulmajid and Christoph Schmid and Jochen J. Frietsch and Helene Schmeller and Tobias A. W. Holderried and Elisa Sala and Helga Schmetzer and Giuliano Filippini Velazquez and Jan Weller},
  journal = {Frontiers in Immunology},
  year = {2026},
  month = oct,
  doi = {10.3389/fimmu.2026.1934726},
  url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1934726/full},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Composition of immune cells in peripheral blood of patients with leukemia in remission after stem cell transplantation (HSCT) with or without graft-versus-host disease (GvHD): a comparative analysis with healthy donors
AU  - Vivien M. Simon
AU  - Cansu Karatas
AU  - Kristina Schreiner
AU  - Anying Huang
AU  - Annalena Frischhut
AU  - Joudi Abdulmajid
AU  - Christoph Schmid
AU  - Jochen J. Frietsch
AU  - Helene Schmeller
AU  - Tobias A. W. Holderried
AU  - Elisa Sala
AU  - Helga Schmetzer
AU  - Giuliano Filippini Velazquez
AU  - Jan Weller
PY  - 2026
DA  - 2026/10/23
JO  - Frontiers in Immunology
DO  - 10.3389/fimmu.2026.1934726
UR  - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1934726/full
AB  - Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is potentially curative for many hematologic malignancies. Clinical outcomes are critically determined by the balance between graft-versus-malignancy effects and graft-versus-host disease (GvM and GvHD, respectively). We hypothesized that integrated immune phenotyping can distinguish immune tolerance from clinically relevant alloreactivity in patients with hematological neoplasia who are in remission after HSCT. These findings may have implications for GvHD screening and monitoring and could support treatment decision-making.  Methods: Multi-color flow cytometry of peripheral blood mononuclear cells profiled myeloid, adaptive, and innate compartments in healthy donors ( n = 10) and post-HSCT patients in remission: without GvHD [NoGvHD ( n = 19)], with chronic (c) GvHD ( n = 18, including n = 4 overlap-a/cGvHD), and with acute (a) GvHD ( n = 6). Frequencies of immune cell subsets were compared between cohorts and stratified by clinical features [e.g., severity, organ involvement, donor lymphocyte infusion (DLI), GvHD development, and cGvHD progression].  Results: NoGvHD patients (clinically asymptomatic and whose disease were in remission) showed immune-cell composition differing substantially from healthy donors: CD14 ⁺ monocytes (Mo), M-MDSCs, and especially checkpoint molecule (cpm)-expressing Mo (TIM3, 2B4, and PDL1) were reduced, while CTLA-4 ⁺ polymorphonuclear myeloid-derived suppressor cells (CTLA-4 ⁺ -PMN-MDSCs) were elevated, as well as T-effector memory (Tem) cells and cpm-expressing T cells (PD1, LAG3, 2B4, TIM3, and TIGIT). cGvHD showed eroded regulatory myeloid subsets and skewed T cells toward effector-memory dominance with increased frequencies of inhibitory cpm-expressing T cells. aGvHD showed marked late-proliferating T-cell expansion (T prol-late /T ³⁺ fold change 9.92 vs. Healthy), T reg depletion, and loss of regulatory NK 56bri populations. Overlap a/cGvHD displayed chronic T-cell stimulation with altered tissue homing (Intβ7 ⁺ enriched), and DLI-induced GvHD showed effector differentiation with insufficient regulatory compensation.  Conclusion: NoGvHD is not “Healthy minus disease” but a distinct post-HSCT immune state. Multi-compartment immune signature profiling by flow cytometry distinguishes preserved tolerance (Healthy) from post-HSCT remodeling (NoGvHD), progressive dysregulation (cGvHD), and acute hyperactivation (aGvHD), and refines GvHD subgroups by clinical context.
SN  - 1664-3224
ER  -