Di-Chimeric Cell Therapy Derived From Hematopoietic and Mesenchymal Stem Cells Promotes Immune Tolerance and Extends Vascularized Composite Allograft Survival

Maria Siemionow; Safak Halil Uygur; Katarzyna Stawarz; Lucile Chambily; Katarzyna Budzynska; Weronika Radecka

doi:10.2478/aite-2026-0013

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Di-Chimeric Cell Therapy Derived From Hematopoietic and Mesenchymal Stem Cells Promotes Immune Tolerance and Extends Vascularized Composite Allograft Survival

Archivum Immunologiae et Therapiae Experimentalis

Volume 74 (2026): Issue 1 (January 2026)

By: Maria Siemionow, Safak Halil Uygur, Katarzyna Stawarz, Lucile Chambily, Katarzyna Budzynska and Weronika Radecka

Open Access

|Apr 2026

Abstract

Chimerism-based strategies remain promising for tolerance induction in solid organ and vascularized composite allograft (VCA) transplantation. This study aimed to develop a novel, less toxic chimeric cell therapy to prolong allograft survival and reduce the need for lifelong immunosuppression. Di-chimeric cells (DCC) were created via polyethylene glycol (PEG)-mediated ex vivo fusion of allogeneic hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) derived from August Copenhagen Irish (ACI) and Lewis rats. Twenty-four fully major histocompatibility complex (MHC)-mismatched groin flap VCAs were transplanted from ACI rat major histocompatibility complex (rat MHC) (RT1^a) donors to Lewis (RT1¹) recipients under a 7-day immunosuppressive protocol of anti-αβTCR antibody and tacrolimus, combined with four different cell therapies of n = 6/group: Group 1, saline control; Group 2, MSC; Group 3, HSC/HSC DCC; and Group 4, HSC/MSC DCC. DCC were delivered via the intraosseous injection. DCC phenotype was confirmed by flow cytometry (FC). Graft rejection was evaluated macroscopically. A single DCC dose significantly prolonged VCA survival, with the best results in Group 4 (94 ± 1.65 days), followed by Group 3 (66 ± 1.24 days), Group 2 (45.5 ± 4.08 days), and Group 1 (38 ± 4.29 days). This study confirmed immunomodulatory and tolerogenic properties of DCC, supporting VCA transplantation.

References

Arslan E, Klimczak A, Siemionow M (2007) Chimerism induction in vascularized bone marrow transplants augmented with bone marrow cells. Microsurgery 27:190–199. https://doi.org/10.1002/micr.20330
Search in Google Scholar Back to article
Bastani S, Staal FJT, Canté-Barrett K (2023) The quest for the holy grail: Overcoming challenges in expanding human hematopoietic stem cells for clinical use. Stem Cell Investig 10:15. https://doi.org/10.21037/sci-2023-016
Search in Google Scholar Back to article
Cohen JL, Trenado A, Vasey D et al. (2002) CD4⁺ CD25⁺ immunoregulatory T cells: New therapeutics for graft-versus-host disease. J Exp Med 196:401–406. https://doi.org/10.1084/jem.20020090
Search in Google Scholar Back to article
Cwykiel J, Jundzill A, Klimczak A et al. (2021a) Donor-recipient chimeric cells induce chimerism and extend survival of vascularized composite allografts. Arch Immunol Ther Exp 69:13. https://doi.org/10.1007/s00005-021-00614-9
Search in Google Scholar Back to article
Cwykiel J, Madajka-Niemeyer M, Siemionow M (2021b) Development of donor–recipient chimeric cells of bone marrow origin as a novel approach for tolerance induction in transplantation. Stem Cell Investig 8:8. https://doi.org/10.21037/sci-2020-044
Search in Google Scholar Back to article
Cyran M, Stawarz K, Chambily L et al. (2024) Assessment of hematopoietic response to total body irradiation in a rat experimental model. Ann Plast Surg 93:100–106. https://doi.org/10.1097/SAP.0000000000003962
Search in Google Scholar Back to article
de Vasconcelos P, Lacerda JF (2022) Hematopoietic stem cell transplantation for neurological disorders: A focus on inborn errors of metabolism. Front Cell Neurosci 16:895511. https://doi.org/10.3389/fncel.2022.895511
Search in Google Scholar Back to article
Doglio M, Crossland RE, Alho AC et al. (2022) Cell-based therapy in prophylaxis and treatment of chronic graft-versus-host disease. Front Immunol 13:1045168. https://doi.org/10.3389/fimmu.2022.1045168
Search in Google Scholar Back to article
Eldaly AS, Avila FR, Torres-Guzman RA et al. (2024) Cell-based therapies induce tolerance of vascularized composite allotransplants: A systematic review. J Surg Res 300:389–401. https://doi.org/10.1016/j.jss.2024.04.079
Search in Google Scholar Back to article
Hall BM (2016) CD4⁺CD25⁺ T regulatory cells in transplantation tolerance: 25 years on. Transplantation 100:2533–2547. https://doi.org/10.1097/TP.0000000000001436
Search in Google Scholar Back to article
Heyes R, Iarocci A, Tchoukalova Y et al. (2016) Immunomodulatory role of mesenchymal stem cell therapy in vascularized composite allotransplantation. J Transplant 2016:6951693. https://doi.org/10.1155/2016/6951693
Search in Google Scholar Back to article
Hivelin M, Klimczak A, Cwykiel J et al. (2016) Immunomodulatory effects of different cellular therapies of bone marrow origin on chimerism induction and maintenance across MHC barriers in a face allotransplantation model. Arch Immunol Ther Exp 64:299–310. https://doi.org/10.1007/s00005-015-0380-8
Search in Google Scholar Back to article
Jundziłł A, Klimczak A, Sonmez E et al. (2021) The positive impact of donor bone marrow cells transplantation into immunoprivileged compartments on the survival of vascularized skin allografts. Arch Immunol Ther Exp 69:28. https://doi.org/10.1007/s00005-021-00631-8
Search in Google Scholar Back to article
Klimczak A, Unal S, Jankowska A et al. (2007) Donor-origin cell engraftment after intraosseous or intravenous bone marrow transplantation in a rat model. Bone Marrow Transplant 40:373–380. https://doi.org/10.1038/sj.bmt.1705743
Search in Google Scholar Back to article
Li Z, Czechowicz A, Scheck A et al. (2019) Hematopoietic chimerism and donor-specific skin allograft tolerance after non-genotoxic CD117 antibody–drug-conjugate conditioning in MHC-mismatched allotransplantation. Nat Commun 10:616. https://doi.org/10.1038/s41467-018-08202-w
Search in Google Scholar Back to article
Ménard C, Tarte K (2013) Immunoregulatory properties of clinical grade mesenchymal stromal cells: Evidence, uncertainties, and clinical application. Stem Cell Res Ther 4:64. https://doi.org/10.1186/scrt214
Search in Google Scholar Back to article
Okeke EB, Uzonna JE (2019) The pivotal role of regulatory T cells in the regulation of innate immune cells. Front Immunol 10:680. https://doi.org/10.3389/fimmu.2019.00680
Search in Google Scholar Back to article
Ou Q, Power R, Griffin MD (2023) Revisiting regulatory T cells as modulators of innate immune response and inflammatory diseases. Front Immunol 14:1287465. https://doi.org/10.3389/fimmu.2023.1287465
Search in Google Scholar Back to article
Ruiz R, Kirk AD (2015) Long-Term Toxicity of Immunosuppressive Therapy. In: Transplantation of the Liver (Third Edition). Philadelphia, PA, USA: W.B. Saunders, pp.1354–1363. https://doi:10.1016/B978-1-4557-0268-8.00097-X
Search in Google Scholar Back to article
Sakaguchi S, Yamaguchi T, Nomura T et al. (2008) Regulatory T cells and immune tolerance. Cell 133:775–787. https://doi.org/10.1016/j.cell.2008.05.009
Search in Google Scholar Back to article
Siemionow M, Klimczak A (2013) Chimerism-based experimental models for tolerance induction in vascularized composite allografts: Cleveland clinic research experience. Clin Dev Immunol 2013:831410. https://doi.org/10.1155/2013/831410
Search in Google Scholar Back to article
Siemionow M, Klimczak A, Unal S (2005) Different routes of donor-derived hematopoietic stem cell transplantation for donor-specific chimerism induction across MHC barrier. Transplant Proc 37:62–64. https://doi.org/10.1016/j.transproceed.2004.12.216
Search in Google Scholar Back to article
Siemionow M, Nasir S (2007) Chimerism and bone marrow-based therapies in transplantation. Microsurgery 27:510–521. https://doi.org/10.1002/micr.20395
Search in Google Scholar Back to article
Siemionow M, Ortak T, Izycki D et al. (2002) Induction of tolerance in composite-tissue allografts. Transplantation 74:1211–1217. https://doi.org/10.1097/00007890-200211150-00002
Search in Google Scholar Back to article
Siemionow M, Rampazzo A, Gharb BB et al. (2016) The reversed paradigm of chimerism induction: Donor conditioning with recipient-derived bone marrow cells as a novel approach for tolerance induction in vascularized composite allotransplantation. Microsurgery 36:676–683. https://doi.org/10.1002/micr.30041
Search in Google Scholar Back to article
Song N, Scholtemeijer M, Shah K (2020) Mesenchymal stem cell immunomodulation: Mechanisms and therapeutic potential. Trends Pharmacol Sci 41:653–664. https://doi.org/10.1016/j.tips.2020.06.009
Search in Google Scholar Back to article
Spaggiari GM, Capobianco A, Abdelrazik H et al. (2008) Mesenchymal stem cells inhibit natural killer-cell proliferation, cytotoxicity, and cytokine production. Blood 111:1327–1333. https://doi.org/10.1182/blood-2007-02-074997
Search in Google Scholar Back to article
Yu S, Lu J (2023) The potential of mesenchymal stem cells to induce immune tolerance to allogeneic transplants. Transpl Immunol 81:101939. https://doi.org/10.1016/j.trim.2023.101939
Search in Google Scholar Back to article
Zhang W, Wang Y, Zhong F et al. (2023) Donor derived hematopoietic stem cell niche transplantation facilitates mixed chimerism mediated donor specific tolerance. Front Immunol 14:1093302. https://doi.org/10.3389/fimmu.2023.1093302
Search in Google Scholar Back to article
Zor F, Karagoz H, Erdemir AT et al. (2016) Reflectance confocal microscopy as a useful diagnostic tool for monitoring skin-containing vascularized composite allograft rejection: A preliminary study in rats. Microsurgery 36:144–151. https://doi.org/10.1002/micr.22419
Search in Google Scholar Back to article

Articles in this issue

DOI: https://doi.org/10.2478/aite-2026-0013 | Journal eISSN: 1661-4917

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Language: English

Submitted on: Dec 19, 2025

Accepted on: Feb 23, 2026

Published on: Apr 9, 2026

Published by: Hirszfeld Institute of Immunology and Experimental Therapy

In partnership with: Paradigm Publishing Services

Publication frequency: 1 issue per year

Keywords:

Chimeric cell therapy,

Chimerism,

Hematopoietic stem cells,

Mesenchymal stem cells,

Transplantation,

Vascularized composite allograft

Related subjects:

Medicine,

Basic medical science,

Biochemistry,

Immunology,

Clinical medicine,

Clinical medicine, other,

Clinical chemistry

© 2026 Maria Siemionow, Safak Halil Uygur, Katarzyna Stawarz, Lucile Chambily, Katarzyna Budzynska, Weronika Radecka, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Volume 74 (2026): Issue 1 (January 2026)