Volume 821 of the series Methods in Molecular Biology pp 279-293


Expanding Human T Regulatory Cells with the mTOR-Inhibitor Rapamycin

  • Manuela BattagliaAffiliated withSan Raffaele Telethon Institute for Gene TherapySan Raffaele Diabetes Research Institute Email author 
  • , Angela StabiliniAffiliated withSan Raffaele Diabetes Research Institute
  • , Eleonora TresoldiAffiliated withSan Raffaele Telethon Institute for Gene Therapy

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CD4+CD25+FOXP3+ T regulatory (Treg) cells are pivotal for the induction and maintenance of peripheral tolerance in both mice and humans. The possibility to use Treg cells for the treatment of T-cell-mediated diseases has recently gained increasing momentum. However, given the limited amount of circulating FOXP3+ Treg cells, efficient methods for their ex vivo expansion are highly desirable. Rapamycin allows for in vitro expansion of murine and human FOXP3+ Treg cells, which maintain their regulatory phenotype and suppressive capacity. Here, we describe in detail the powerful methods for enriching human FOXP3+ Treg cells starting from unfractionated CD4+ T cells or for expanding CD25+-enriched Treg cells in the presence of rapamycin.

Key words

Immunological tolerance Regulatory T cells Rapamycin