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@ARTICLE{Lakshmi:307321,
      author       = {K. Lakshmi and A. v. Jutrzenka-Trzebiatowski and L.
                      Loureiro and K. E. G. Soto and K. Peter and J. M. M. Morales
                      and S. S. Nirmala and N. Berndt and C. Arndt and Y. Hu and
                      J.-W. Li and C. Peitzsch and A. Taubenberger and R.
                      Wehner$^*$ and M. Schmitz$^*$ and K. Hölig and H. Abken and
                      E. Bonifacio and M. Bornhäuser$^*$ and M. Bachmann$^*$ and
                      A. Feldmann$^*$ and A. Fuchs$^*$},
      title        = {{A}utomated {GMP}-compatible production of universal {CAR}
                      {T}regs for organ-targeted tolerance induction.},
      journal      = {Journal of translational medicine},
      volume       = {23},
      number       = {1},
      issn         = {1479-5876},
      address      = {London},
      publisher    = {BioMed Central},
      reportid     = {DKFZ-2025-03007},
      pages        = {1399},
      year         = {2025},
      abstract     = {Adoptive transfer of regulatory T cells (Tregs) has
                      demonstrated safety, feasibility and early signs of efficacy
                      in promoting immunological tolerance in inflammatory
                      conditions such as graft-versus-host disease (GvHD).
                      Chimeric antigen receptor (CAR)-engineered Tregs offer
                      localized activation and suppression compared to polyclonal
                      Tregs, but their clinical translation is limited by high
                      manufacturing costs, lengthy developing times and fixed
                      single-antigen specificity. To address these limitations, we
                      employed the universal adapter Reverse CAR (RevCAR) system,
                      which harbors a peptide epitope lacking intrinsic antigen
                      specificity but provides flexibility in targeting through
                      the use of an antigen-specific RevCAR Target Module (RevTM).
                      As a proof-of-concept, we used a RevTM targeting
                      carcinoembryonic antigen (CEA), which is highly expressed in
                      the gastrointestinal (GI) tract, as a potential strategy to
                      achieve localized immunosuppression in GI acute GvHD.To
                      support clinical translation, we established an automated,
                      GMP-compatible, clinical-scale manufacturing process. Tregs
                      were magnetically enriched from leukapheresis using the
                      CliniMACS® Plus, followed by high-purity sorting on the
                      MACSQuant® Tyto®. The sorted cells were virally transduced
                      and the RevCAR Tregs were expanded on the CliniMACS
                      Prodigy® to obtain clinically relevant cell numbers. The
                      harvested products were evaluated for phenotype, stability,
                      antigen specificity and suppressive function.Across five
                      manufacturing runs, Tregs (CD4+CD25highCD127lowFOXP3+) with
                      a median initial purity of $94\%$ were expanded to achieve a
                      median therapeutic yield of 602 × 106 cells. The final
                      product maintained a high purity (median: $91.9\%)$ and
                      exhibited high RevCAR expression (median: $60\%$ RevCAR+).
                      Mass cytometry analysis revealed that expanded RevCAR Tregs
                      predominantly exhibited a central memory phenotype with high
                      expression of functional and homing markers. Under
                      experimental pro-inflammatory conditions, the cells
                      maintained stable FOXP3 and Helios expression with minimal
                      pro-inflammatory cytokine production. Importantly, RevCAR
                      Tregs showed antigen-specific activation upon target
                      engagement via the CEA-specific RevTM and robust,
                      dose-dependent suppression.The study establishes a scalable,
                      GMP-compatible process for manufacturing pure, stable and
                      functional universal RevCAR Tregs for clinical applications.
                      Furthermore, the RevCAR system offers a promising approach
                      toward an allogenic, off-the-shelf Treg therapy capable of
                      treating diverse immune-mediated diseases with spatial
                      precision.},
      keywords     = {T-Lymphocytes, Regulatory: immunology / T-Lymphocytes,
                      Regulatory: cytology / Humans / Receptors, Chimeric Antigen:
                      metabolism / Immune Tolerance: immunology / Automation /
                      Organ Specificity / Carcinoembryonic Antigen: metabolism /
                      Carcinoembryonic Antigen: immunology / Graft vs Host
                      Disease: immunology / Automated expansion (Other) /
                      Closed-system manufacturing (Other) / Good manufacturing
                      practice (Other) / Precision immunotherapy (Other) /
                      Regulatory T cells (Other) / Treg cell therapy (Other) /
                      Universal adapter CAR (Other) / Receptors, Chimeric Antigen
                      (NLM Chemicals) / Carcinoembryonic Antigen (NLM Chemicals)},
      cin          = {DD01},
      ddc          = {610},
      cid          = {I:(DE-He78)DD01-20160331},
      pnm          = {899 - ohne Topic (POF4-899)},
      pid          = {G:(DE-HGF)POF4-899},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:41408297},
      doi          = {10.1186/s12967-025-07431-0},
      url          = {https://inrepo02.dkfz.de/record/307321},
}