Column-free isolation of untouched CD4+CD127lowCD49d- human regulatory T cells in 45 minutes (168.4)

Abstract

Abstract Regulatory T cells (Tregs) are a subset of lymphocytes that play a key role in maintaining immune homeostasis. The isolation of highly purified Tregs is essential for advancing this exciting field of research. FOXP3 remains the best marker for the identification of Tregs, but its intracellular localization currently precludes its use for the isolation of viable human Tregs. To overcome these technical difficulties, researchers have exploited the expression of other surface markers to isolate Treg populations. Typically, the expression of CD127 inversely correlates with FOXP3 expression, while CD49d, is expressed on the majority of pro-inflammatory effector cells but is absent on Tregs. Removal of CD127high cells from the CD4+ T-cell population enriches for Tregs with high levels of FOXP3 expression. Additional depletion of CD49d expressing cells further removes contaminating IFNγ and IL-17 secreting cells. Together this strategy permits selective isolation of highly enriched Tregs. We have developed a new kit for the negative enrichment of highly purified human Tregs. Our new EasySep™ CD4+CD127lowCD49d- kit isolates untouched Tregs by by column-free magnetic separation in only 45 minutes. Mean CD4+ T-cell purity is 94.3% +/- 5.5% of which 65.5% +/- 11.7% express high levels of FOXP3 (n=7). This new product will add to the array of powerful yet convenient tools available for the isolation of functional highly purified human Tregs.