Low Dose of IL-2 Application for Graft-Versus-Host Disease Prophylaxis Following Haploidentical Stem Cell Transplantation

Abstract

Graft-versus-host disease (GVHD) are still key obstacles of haploidentical transplantation. Interleukin-2 (IL-2) could promote natural killer (NK) cells and T-regulatory cells (Tregs) cells expansion in vitro and in vivo. We explored whether low-dose IL-2 administration at an early stage could promote NK cells and Tregs reconstitution and reduce GVHD after haplo-HSCT. This cohort trial included 10 recipients of accepting IL-2 treatment and case-pairing 30 recipients without IL-2 treatment post haplo-HSCT. In contrast to the control group, the 5-year incidence of chronic GVHD (cGVHD) was lower (p = 0.018), and GVHD progression-free survival (GPFS) was better (p = 0.025) in the IL-2 group. Blood NK-cells, Treg cells, conventional T cells (Tcon) cells, and the expression of CD62L+ on Tregs and Tcon cells reconstitution were increased post-IL-2 treatment. NKG2A expression on NK cells increased significantly post-IL-2 treatment. Meanwhile, IL-2 administration shortly increased the plasma levels of IFN-Ƴ, TNF-a, IL-10, and IL-2 in subjects post haplo-HSCT. Relative to the control group, low-dose IL-2 increased NK cell counts and the expression of CD122, DNAM-1, and NKG2D on NK cells post transplantation. Administration of low-dose IL-2 after haplo-HSCT correlated with reduced cGVHD, which should be explored further with randomized trial.

Keywords

graft-versus-host disease / haploidentical transplantation / interleukin-2 / natural killer cells / T-regulatory cells

References

[1] O. Penack, M. Marchetti, M. Aljurf, et al., “Prophylaxis and Management of Graft-Versus-Host Disease After Stem-Cell Transplantation for Haematological Malignancies: Updated Consensus Recommendations of the European Society for Blood and Marrow Transplantation,” Lancet Haematology 11, no. 2 (2024): e147–e159.

[2] S. Naik, Y. Li, A. C. Talleur, et al., “Memory T-Cell Enriched Haploidentical Transplantation With NK Cell Addback Results in Promising Long-term Outcomes: A Phase II Trial,” Journal of Hematology & Oncology 17, no. 1 (2024): 50.

[3] V. D. Kheav, M. Busson, C. Scieux, et al., “Favorable Impact of Natural Killer Cell Reconstitution on Chronic Graft-Versus-Host Disease and Cytomegalovirus Reactivation After Allogeneic Hematopoietic Stem Cell Transplantation,” Haematologica 99, no. 12 (2014): 1860–1867.

[4] L. Ruggeri, M. Capanni, E. Urbani, et al., “Effectiveness of Donor Natural Killer Cell Alloreactivity in Mismatched Hematopoietic Transplants,” Science 295, no. 5562 (2002): 2097–2100.

[5] A. Zafarani, M. Taghavi-Farahabadi, M. H. Razizadeh, M. R. Amirzargar, M. Mansouri, and M. Mahmoudi, “The Role of NK Cells and Their Exosomes in Graft Versus Host Disease and Graft Versus Leukemia,” Stem Cell Reviews and Reports 19, no. 1 (2023): 26–45.

[6] M. Edinger, P. Hoffmann, J. Ermann, et al., “CD4+CD25+ Regulatory T Cells Preserve Graft-Versus-Tumor Activity While Inhibiting Graft-Versus-Host Disease After Bone Marrow Transplantation,” Nature Medicine 9, no. 9 (2003): 1144–1150.

[7] S. Ito, C. M. Bollard, M. Carlsten, et al., “Ultra-Low Dose Interleukin-2 Promotes Immune-Modulating Function of Regulatory T Cells and Natural Killer Cells in Healthy Volunteers,” Molecular Therapy: The Journal of the American Society of Gene Therapy 22, no. 7 (2014): 1388–1395.

[8] O. Boyman and J. Sprent, “The Role of Interleukin-2 During Homeostasis and Activation of the Immune System,” Nature Reviews Immunology 12, no. 3 (2012): 180–190.

[9] A. I. Palamarchuk, E. I. Kovalenko, and M. A. Streltsova, “The hTERT and iCasp9 Transgenes Affect EOMES and T-BET Levels in NK Cells and the Introduction of both Genes Improves NK Cell Proliferation in Response to IL2 and IL15 Stimulation,” Biomedicines 12, no. 3 (2024): 650.

[10] V. Niederlova, O. Tsyklauri, M. Kovar, and O. Stepanek, “IL-2-driven CD8(+) T Cell Phenotypes: Implications for Immunotherapy,” Trends in Immunology 44, no. 11 (2023): 890–901.

[11] B. C. Betts, J. Pidala, J. Kim, et al., “IL-2 Promotes Early Treg Reconstitution After Allogeneic Hematopoietic Cell Transplantation,” Haematologica 102, no. 5 (2017): 948–957.

[12] A. A. Kennedy-Nasser, S. Ku, P. Castillo-Caro, et al., “Ultra Low-Dose IL-2 for GVHD Prophylaxis After Allogeneic Hematopoietic Stem Cell Transplantation Mediates Expansion of Regulatory T Cells Without Diminishing Antiviral and Antileukemic Activity,” Clinical Cancer Research 20, no. 8 (2014): 2215–2225.

[13] X. Y. Zhao, X. S. Zhao, Y. T. Wang, et al., “Prophylactic Use of Low-Dose Interleukin-2 and the Clinical Outcomes of Hematopoietic Stem Cell Transplantation: A Randomized Study,” Oncoimmunology 5, no. 12 (2016): e1250992.

[14] D. Wolf, “GVHD Prophylaxis: Use an Ortho IL-2/IL-2Rbeta Treg System,” Blood 141, no. 11 (2023): 1246–1247.

[15] X. D. Ma, Z. L. Xu, and X. J. Huang, “Immune Reconstitution After Haploidentical Hematopoietic Stem Cell Transplantation With Different Non-T-Cell Depletion Protocols,” MedComm 6, no. 6 (2025): e70206.

[16] J. Koreth, K. Matsuoka, H. T. Kim, et al., “Interleukin-2 and Regulatory T Cells in Graft-Versus-Host Disease,” New England Journal of Medicine 365, no. 22 (2011): 2055–2066.

[17] K. Matsuoka, H. T. Kim, S. McDonough, et al., “Altered Regulatory T Cell Homeostasis in Patients With CD4+ Lymphopenia Following Allogeneic Hematopoietic Stem Cell Transplantation,” The Journal of Clinical Investigation 120, no. 5 (2010): 1479–1493.

[18] J. Koreth, H. T. Kim, K. T. Jones, et al., “Efficacy, Durability, and Response Predictors of Low-Dose Interleukin-2 Therapy for Chronic Graft-Versus-Host Disease,” Blood 128 (2016): 130–137.

[19] J. S. Whangbo, S. Nikiforow, H. T. Kim, et al., “A Phase 1 Study of Donor Regulatory T-cell Infusion plus Low-dose Interleukin-2 for Steroid-refractory Chronic Graft-vs-host Disease,” Blood Advances 6, no. 21 (2022): 5786–5796.

[20] T. Meyer, K. Maas-Bauer, R. Wasch, et al., “Immunological Reconstitution and Infections After alloHCT—A Comparison Between Post-transplantation Cyclophosphamide, ATLG and Non-ATLG Based GvHD Prophylaxis,” Bone Marrow Transplantation 60, no. 3 (2025): 286–296.

[21] B. Bielekova, M. Catalfamo, S. Reichert-Scrivner, et al., “Regulatory CD56(Bright) Natural Killer Cells Mediate Immunomodulatory Effects of IL-2Ralpha-Targeted Therapy (Daclizumab) in Multiple Sclerosis,” Proceedings of the National Academy of Sciences of the United States of America 103, no. 15 (2006): 5941–5946.

[22] V. D. Kheav, M. Busson, C. Scieux, et al., “Favorable Impact of Natural Killer Cell Reconstitution on Chronic Graft-Versus-Host Disease and Cytomegalovirus Reactivation After Allogeneic Hematopoietic Stem Cell Transplantation,” Haematologica 99, no. 12 (2014): 1860–1867.

[23] T. A. Fehniger, M. A. Cooper, G. J. Nuovo, et al., “CD56bright Natural Killer Cells Are Present in Human Lymph Nodes and Are Activated by T Cell-Derived IL-2: A Potential New Link Between Adaptive and Innate Immunity,” Blood 101, no. 8 (2003):3052–3057.

[24] R. S. Goldszmid, P. Caspar, A. Rivollier, et al., “NK Cell-derived Interferon-Gamma Orchestrates Cellular Dynamics and the Differentiation of Monocytes Into Dendritic Cells at the Site of Infection,” Immunity 36, no. 6 (2012): 1047–1059.

[25] Y. J. Chang, X. Y. Zhao, and X. J. Huang, “Effects of the NK Cell Recovery on Outcomes of Unmanipulated Haploidentical Blood and Marrow Transplantation for Patients With Hematologic Malignancies,” Biology of Blood and Marrow Transplantation 14, no. 3 (2008): 323–334.

[26] K. Umino, K. Morita, T. Ikeda, et al., “Antibody-Mediated Pathogenesis of Chronic GVHD Through DBY/HLA Class II Complexes and Induction of a GVL Effect,” Blood 142, no. 11 (2023): 1008–1021.

[27] M. H. Sofi, L. Tian, S. Schutt, et al., “Ceramide Synthase 6 Impacts T-Cell Allogeneic Response and Graft-Versus-Host Disease Through Regulating N-RAS/ERK Pathway,” Leukemia 36, no. 7 (2022):1907–1915.

[28] W. H. Hallett, E. Ames, M. Alvarez, et al., “Combination Therapy Using IL-2 and Anti-CD25 Results in Augmented Natural Killer Cell-mediated Antitumor Responses,” Biology of Blood and Marrow Transplantation: Journal of the American Society for Blood and Marrow Transplantation 14, no. 10 (2008): 1088–1099.

[29] V. Bachanova, S. Cooley, T. E. Defor, et al., “Clearance of Acute Myeloid Leukemia by Haploidentical Natural Killer Cells Is Improved Using IL-2 Diphtheria Toxin Fusion Protein,” Blood 123, no. 25 (2014): 3855–3863.

[30] M. R. Verneris, Natural Killer Cells and Regulatory T Cells: How to Manipulate a Graft for Optimal GVL (ASH Education Program Book, 2013).

[31] L. Q. Cao, W. X. Huo, X. H. Zhang, et al., “Peripheral Blood Stem Cell Transplantation From Haploidentical Related Donor Could Achieve Satisfactory Clinical Outcomes for Intermediate- or High-Risk Adult Acute Myeloid Leukemia Patients,” Bone Marrow Transplantation 59, no. 2 (2024): 203–210.

[32] Q. Shang, L. Bai, Y. Cheng, et al., “Outcomes and Prognosis of Haploidentical Haematopoietic Stem Cell Transplantation in Children With FLT3-ITD Mutated Acute Myeloid Leukaemia,” Bone Marrow Transplantation 59, no. 6 (2024): 824–831.

[33] Z. L. Xu, T. T. Han, X. L. Zhu, et al., “Randomized Trial of Anti-Thymocyte Globulin Plus Lowdose Post-transplant Cyclophosphamide to Prevent Graft-Versus-Host Disease in Haploidentical Transplantation,” Haematologica 110, no. 12 (2025): 2965–2973.

[34] Z. Xu, X. Mo, Y. Kong, et al., “Mini-Dose Methotrexate Combined With Methylprednisolone as a First-Line Treatment for Acute Graft-Versus-Host Disease: A Phase 2 Trial,” Journal of Translational Internal Medicine 11, no. 3 (2023): 255–264.

[35] Y. Wang, Q. F. Liu, D. P. Wu, et al., “Mini-Dose Methotrexate Combined With Methylprednisolone for the Initial Treatment of Acute GVHD: A Multicentre, Randomized Trial,” BMC Medicine 22, no. 1 (2024): 176.

[36] X. Y. Pei, Q. Huang, L. J. Luo, et al., “Letermovir Prophylaxis for Cytomegalovirus Is Associated With Risk of Post-Transplant Lymphoproliferative Disorders After Haploidentical Stem Cell Transplantation,” Haematologica 110, no. 4 (2025): 1005–1009.

[37] O. Penack, M. Marchetti, M. Aljurf, et al., “Prophylaxis and Management of Graft-Versus-Host Disease After Stem-Cell Transplantation for Haematological Malignancies: Updated Consensus Recommendations of the European Society for Blood and Marrow Transplantation,” Lancet Haematology 11, no. 2 (2024): e147–e159.

[38] M. H. Jagasia, H. T. Greinix, M. Arora, et al., “National Institutes of Health Consensus Development Project on Criteria for Clinical Trials in Chronic Graft-Versus-Host Disease: I. The 2014 Diagnosis and Staging Working Group Report,” Biology of Blood and Marrow Transplantation 21, no. 3 (2015): 389–401. e381.

RIGHTS & PERMISSIONS

2026 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.