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
2012 Immunology and Cell Biology  
Inhibition of activation induced CD154 on CD4⁺ CD25⁻ cells: a valid surrogate for human Treg suppressor function, Immunology and Cell Biology, 2012; 90(8):812-821. Abstract Natural Regulatory T cells (Treg) 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 homeostasis is the subject of ongoing
more » ... , 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 leukocyte reaction (MLR) over a 3-6 day time period. As an alternative to this assay we show that a 7 hr CD154 expression assay is rapid, simple, and provides a reliable readout of suppressor function. Using multiple Treg-like cell types including nTreg, iTreg 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.
doi:10.1038/icb.2012.18 pmid:22689014 fatcat:xozdwmrw3rg5xiekfktxk7dq2i