OMIP‐053: Identification, Classification, and Isolation of Major FoxP3 Expressing Human CD4⁺ Treg Subsets

Abstract

WE designed and optimized an inter-convertible flow cytometry panel for the analysis and sorting of human CD4+ regulatory T cells (Treg) utilizing all of the major, currently accepted marker combinations for Treg identification. The panel is optimized for use with peripheral blood mononuclear cells (PBMC). In addition, the panel allows for the identification, classification, and isolation of activated, antigen-experienced Treg, and provides estimates of proliferation and suppressive capacity. To this end we created two, easily inter-convertible sub-panels: ATREG—which includes intra-nuclear markers, and BTREG—using surface markers only. Both sub-panels have been tested on cryopreserved PBMC from healthy donors and those with human autoimmune diseases, as well as on long-term cultured human Treg and non-Treg cell lines and clones.