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De novo production of antigen-specific suppressor cells in vivo

Nature Protocols volume 1, pages 653–661 (2006)Cite this article

Abstract

Foxp3-expressing regulatory T cells (Treg) play an essential role in maintaining tolerance to self antigens and are generated under physiological conditions when developing T cells encounter antigens expressed by thymic epithelial cells. We have addressed the possibility that Treg can be exploited to prevent or even suppress ongoing immune responses to foreign antigens. To this end, one must develop methods that permit the de novo generation of Treg specific for foreign antigens in peripheral lymphoid tissue. This report describes the methodology of generating Treg by delivering minute doses of peptide contained in fusion Abs directed against the DEC-205 endocytic receptor on steady-state dendritic cells. The process, from cloning and production of fusion Abs to antigen-specific Treg induction in vivo, takes ∼2 months. The results show that delivery of T-cell receptor agonist ligands under subimmunogenic conditions represents a suitable approach for converting naive T cells into Treg.

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Figure 1: Design, dimerization and cloning of oligonucleotides encoding the peptide of interest with spacing residues.
Figure 2: Analysis of oligonucleotide dimerization.
Figure 3: Anti-DEC-HA107–119 delivery of antigen to DCs in vitro.
Figure 4: Conversion of naive Foxp3−CD4+CD25− T cells into Foxp3+CD4+CD25+ T cells in vivo.

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Acknowledgements

We would like to thank L. Benson for help in preparing the manuscript.

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Authors and Affiliations

  1. Department of Pathology, Harvard Medical School, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Karsten Kretschmer, Tracy S P Heng & Harald von Boehmer

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  1. Karsten Kretschmer
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  2. Tracy S P Heng
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  3. Harald von Boehmer
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Corresponding author

Correspondence to Harald von Boehmer.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Complete nucleotide sequence coding for the anti-DEC-205 Ig heavy chain fused to the HA107–119 peptide. (PDF 2669 kb)

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Kretschmer, K., Heng, T. & von Boehmer, H. De novo production of antigen-specific suppressor cells in vivo. Nat Protoc 1, 653–661 (2006). https://doi.org/10.1038/nprot.2006.105

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