Differentiation of Peripheral Treg

Methods Mol Biol. 2023:2559:67-77. doi: 10.1007/978-1-0716-2647-4_6.

Abstract

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 transfer; DEREG; DO11.10; Foxp3; Oral tolerance; Treg; pTreg differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Diphtheria Toxin
  • Drinking Water*
  • Forkhead Transcription Factors
  • Heparin-binding EGF-like Growth Factor
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Ovalbumin
  • T-Lymphocytes, Regulatory*

Substances

  • Diphtheria Toxin
  • Drinking Water
  • Forkhead Transcription Factors
  • Heparin-binding EGF-like Growth Factor
  • Ovalbumin