Antigen-reactive regulatory T cells can be expanded in vitro with monocytes and anti-CD28 and anti-CD154 antibodies

Research output: Contribution to journalResearch articleContributedpeer-review


  • Dorota Iwaszkiewicz-Grzes - , Medical University of Gdańsk, Poltreg S.A. (Author)
  • Mateusz Gliwinski - , Medical University of Gdańsk, Poltreg S.A. (Author)
  • Anne Eugster - , TUD Dresden University of Technology (Author)
  • Magdalena Piotrowska - , Medical University of Gdańsk (Author)
  • Andreas Dahl - , DRESDEN-concept Genome Center (CMCB Core Facility) (Author)
  • Natalia Marek-Trzonkowska - , Medical University of Gdańsk, University of Gdańsk, Poltreg S.A. (Author)
  • Piotr Trzonkowski - , Medical University of Gdańsk, Poltreg S.A. (Author)


Background: In recent years, therapies with CD4+CD25highFoxP3+ regulatory T cells (Tregs) have been successfully tested in many clinical trials. The important issue regarding the use of this treatment in autoimmune conditions remains the specificity toward particular antigen, as because of epitope spread, there are usually multiple causative autoantigens to be regulated in such conditions. Methods: Here we show a method of generation of Tregs enriched with antigen-reactive clones that potentially covers the majority of such autoantigens. In our research, Tregs were expanded with anti-CD28 and anti-CD154 antibodies and autologous monocytes and loaded with a model peptide, such as whole insulin or insulin β chain peptide 9–23. The cells were then sorted into cells recognizing the presented antigen. The reactivity was verified with functional assays in which Tregs suppressed proliferation or interferon gamma production of autologous effector T cells (polyclonal and antigen-specific) used as responders challenged with the model peptide. Finally, we analyzed clonotype distribution and TRAV gene usage in the specific Tregs. Results: Altogether, the applied technique had a good yield and allowed us to obtain a Treg product enriched with a specific subset, as confirmed in the functional tests. The product consisted of many clones; nevertheless, the content of these clones was different from that found in polyclonal or unspecific Tregs. Conclusions: The presented technique might be used to generate populations of Tregs enriched with cells reactive to any given peptide, which can be used as a cellular therapy medicinal product in antigen-targeted therapies.


Original languageEnglish
Pages (from-to)629-641
Number of pages13
Issue number11
Publication statusPublished - Nov 2020

External IDs

PubMed 32778404


Sustainable Development Goals


  • antigen stimulation, antigen-reactive, antigen-specific, immune therapies, regulatory T cells

Library keywords