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Updated: Aug 27, 2025

Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells
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Differentiation of Peripheral Treg

José Almeida-Santos1,2, Marie-Louise Bergman3, Jocelyne Demengeot3

  • 1Instituto Gulbenkian de Ciência, Oeiras, Portugal. j.almeida-santos@imperial.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2022
PubMed

This chapter shows protocols for the differentiation of peripheral Treg (pTreg) from polyclonal and monoclonal CD4+ T cells. Polyclonal naïve CD4+ T cells can differentiate into pTreg upon adoptive transfer into Foxp3-diphtheria toxin receptor transgenic recipient mice in which endogenous Tregs are transiently depleted by administration of diphtheria toxin before adoptive transfer. Differentiation of monoclonal pTreg is induced through oral delivery of ovalbumin into RAG-deficient DO11.10 mice, in which T cells are ovalbumin specific. We show the isolation of naïve CD4+ T cells by flow cytometry, the administration of ovalbumin in drinking water, and the analysis tools, including an optional protocol for the enrichment of analysis samples in CD4+ T cells using a magnetic purification.

Keywords:
Adoptive T cell transferDEREGDO11.10Foxp3Oral toleranceTregpTreg differentiation

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