HLA matching or CRISPR editing of HLA class I/II enables engraftment and effective function of allogeneic human regulatory T cell therapy in a humanized mouse transplantation model
Regulatory T cells (Tregs) hold promise for treating autoimmune disease and transplant rejection, yet generation of autologous products for adoptive transfer can suffer donor variability and slow turnaround, limiting their use in urgent indications. We therefore examine whether allogeneic, pre-manufactured ('off-the-shelf') Tregs could overcome these barriers. In a human skin-xenograft model, HLA-mismatched Tregs are swiftly eliminated by recipient CD8<sup>+</sup> T cells and fail to protect grafts. Stringent matching of HLA class I and II restores efficacy but is clinically impractical. Using non-viral CRISPR editing we disrupt B2M and CIITA while inserting an HLA-E-B2M fusion, generating hypo-immunogenic Tregs that evade both T and NK cell attack. Engineered cells retain FOXP3 stability and potent in vitro suppression, and after a single low-dose infusion, prolong human skin graft survival in a humanized mouse model comparably to autologous Tregs. Histology and spatial transcriptomics reveal minimal cytotoxic infiltration and enrichment of immunoregulatory and tissue-repair programmes. Multiplex HLA engineering thus enables ready-to-use allogeneic Tregs that withstand host immune attack for adoptive transfer.
Authors
- Dimitrios L. Wagner (ORCID: https://orcid.org/0000-0002-2189-3579)
- Clemens Franke (ORCID: https://orcid.org/0009-0009-0476-2807)
- Helen Stark (ORCID: https://orcid.org/0000-0002-9360-9864)
- Annette Künkele (ORCID: https://orcid.org/0000-0002-8406-5412)
- Sarah Short (ORCID: https://orcid.org/0000-0002-7594-7915)
- М. Yu. Valkov (ORCID: https://orcid.org/0000-0003-3230-9638)
- Julia K. Polansky (ORCID: https://orcid.org/0000-0003-4727-2540)
- Amy Cross (ORCID: https://orcid.org/0000-0002-2592-6251)
- Merve Bilici
- Hans‐Dieter Volk (ORCID: https://orcid.org/0000-0002-7743-6668)
- Fadi Issa (ORCID: https://orcid.org/0000-0002-8279-7732)
- Viktor Glaser (ORCID: https://orcid.org/0000-0002-1385-5249)
- Joanna Hester (ORCID: https://orcid.org/0000-0002-7466-3849)
- Jonas Kath (ORCID: https://orcid.org/0000-0001-7721-7978)
- Oliver McCallion (ORCID: https://orcid.org/0000-0002-5977-812X)
- Petra Reinke (ORCID: https://orcid.org/0000-0003-0771-4375)
- Leila Amini (ORCID: https://orcid.org/0000-0003-4855-5905)
- Michael Schmueck‐Henneresse (ORCID: https://orcid.org/0000-0001-5964-9179)
- Weijie Du (ORCID: https://orcid.org/0000-0001-5924-8631)
- Kate Milward (ORCID: https://orcid.org/0000-0003-3019-4443)
- Mingxing Yang
Institutions
- Baylor College of Medicine (US)
- Humboldt-Universität zu Berlin (DE)
- University of Oxford (GB)
- German Cancer Society (DE)
- Berlin Institute of Health at Charité - Universitätsmedizin Berlin (DE)
- Oxford BioMedica (United Kingdom) (GB)
- Children's Cancer Center (US)
- Deutsches Konsortium für Translationale Krebsforschung (DE)
- Freie Universität Berlin (DE)
- Charité - Universitätsmedizin Berlin (DE)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-10-13
- DOI
- https://doi.org/10.1038/s41467-025-64945-3
- Citations
- 4
- Primary Topic
- CAR-T cell therapy research
- Type
- article
- Field-Weighted Citation Impact
- 4.18
Funders
- British Heart Foundation
- Kidney Research UK
- European Commission
- Chinese Academy of Medical Sciences
- Berlin Institute of Health
- Medical Research Council