Inhibition of activation induced CD154 on CD4 CD25 cells: A valid surrogate for human Treg suppressor function

Danika Hill, Nicola Eastaff-Leung, Suzanne Bresatz-Atkins, Noel Warner, Joyce Ruitenberg, Doreen Krumbiegel, Steve Pederson, Natasha Mcinnes, Cheryl Y. Brown, Timothy Sadlon, Simon C. Barry

Research output: Contribution to journalArticleResearchpeer-review

12 Citations (Scopus)

Abstract

Natural Regulatory T cells (Tregs) are defined by stable expression of the cell surface proteins CD4 and CD25, low surface expression of CD127 and expression of the transcription factor FOXP3. The contribution of Treg to the prevention of autoimmunity and the maintenance of immune homoestasis is the subject of ongoing interest, as alterations in Treg numbers and function are implicated in a wide range of diseases. The in vitro benchmark for determining Treg function is suppression of proliferation of unmatched effector T cells in a mixed lymphocyte reaction (MLR) over a 3-6-day time period. As an alternative to this assay, we show that a 7-h CD154 expression assay is rapid, simple and provides a reliable readout of suppressor function. Using multiple Treg-like cell types including natural (n)Treg, inducible (i)Treg and Treg cell lines, we show that suppression of CD154 expression is a surrogate for suppression of proliferation. We propose this as a suitable alternative to the MLR assay, as it is rapid and may be more amenable to high-throughput screening, analysing large cohorts of clinical samples or assaying transiently suppressive populations.

Original languageEnglish
Pages (from-to)812-821
Number of pages10
JournalImmunology and Cell Biology
Volume90
Issue number8
DOIs
Publication statusPublished - 1 Sep 2012
Externally publishedYes

Keywords

  • functional assays
  • iTreg
  • nTreg
  • regulatory T cells

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