Methods for In Vitro Generation of Human Type 1 Regulatory T Cells

  • Silvia Gregori
  • Maria Grazia Roncarolo
  • Rosa BacchettaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 677)


Type 1 regulatory T (Tr1) cells are adaptive regulatory T cells that are induced in the periphery upon chronic exposure to antigen (Ag) in a tolerogenic environment containing interleukin (IL)-10. Tr1 cells are Ag-specific; they produce high levels of IL-10 and TGF-β in the absence of IL-4 and suppress T-cell responses via a cytokine-dependent mechanism. During the last decade, several protocols have been developed to generate Tr1 cell lines in vitro. In this chapter, we outline protocols to generate non-Ag- and Ag-specific Tr1 cell lines and assays used to characterize Tr1 cell phenotype and functions.

Key words

Type 1 regulatory T cells Dendritic cells Interleukin-10 



The authors thank Claudia Sartirana and Chiara Francesca Magnani for technical help. This work was supported by a grant from the Italian Telethon Foundation, Cariplo Foundation, the Italian Association for Cancer Research (Associazione Italiana per la Ricerca sul Cancro, AIRC), and the EU graut to MGR, Riset Consortion (


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Copyright information

© Humana Press 2010

Authors and Affiliations

  • Silvia Gregori
    • 1
  • Maria Grazia Roncarolo
    • 1
    • 2
  • Rosa Bacchetta
    • 1
    Email author
  1. 1.Department of Regenerative Medicine, Stem Cells, and Gene TherapySan Raffaele Telethon Institute for Gene Therapy (HSR-TIGET)MilanItaly
  2. 2.Universita’ Vita-Salute San RaffaeleMilanItaly

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