Using flow cytometry to first gate on and sort viable cells with markers consistent with Tregs, then functionally testing to see if, as a group, the cells defined by your gating strategy actually act like Tregs, is currently the best way to quantify Tregs in your sample.

In conclusion, we propose an essential marker set comprising antibodies to CD3, CD4, CD25, CD127, Foxp3, Ki67, and CD45RA and a corresponding robust gating strategy for the context-dependent analysis of Tregs by flow cytometry.

Regulatory T (Treg) cells are a subset of CD4 + T cells that is involved in maintaining immune homeostasis and self-tolerance by inhibiting the pro-inflammatory activities of CD4 + and CD8 + effector T cells, natural killer cells, and antigen-presenting cells.

In this study, we describe an optimized multi-parameter flow cytometry protocol for quantification of human Treg cells from freshly obtained and viably frozen samples and correlations with epigenetic Treg cell analysis (TSDR demethylation).

Comprehensive flow cytometry panels for identifying Tregs should include markers associated with immunosuppressive function. We developed, optimized, and validated an 11-color antibody panel for the standardized detection of Tregs from cryopreserved human PBMCs.

We have outlined below a flow cytometric technique to detect and isolate the entire human Treg population based on expression of CD4, CD25, and CD127. Use of a number of additional antibodies for defining subsets within the Treg compartment is described.

Sep 15, 2019 · This chapter describes how to measure human Tregs using flow cytometry for the purposes of immunophenotyping. Flow cytometry requires samples to be in a single cell suspension in order to establish a laminar sample flow through the point of acquisition.

摘要:CD4+Foxp3+ 调节性T细胞 (Treg) 在控制自身免疫耐受和炎症反应方面发挥相当关键的作用 (Sakaguchi,2004) 。 研究表明,T细胞受体 (TCR) 信号控制Treg细胞的活化、维持和免疫抑制功能,并进而影响Treg细胞的基因表达、代谢、粘附和迁移以控制机体的免疫稳态和平衡 (Li 和Rudensky,2016) 。 因此,研究Treg细胞的活化和免疫抑制功能具有重要的意义。 在这里,基于运用荧光染料 ( 如CellTrace Violet、CFSE 等) 标记T细胞,并在每个细胞分裂时在子细胞之间平均 …

Jan 19, 2023 · This study determined the expression of surface markers on human peripheral Treg cells and conventional T (Tconv) cells in a steady state and after activation to gain insight into their mechanism of action and more precisely characterize this regulatory population in …

Regulatory T cells (Tregs) are a critical subset of CD4+ T cells that regulate the immune response and prevent autoimmunity. They are capable of inducing apoptosis, increasing cellular adenosine, and inhibiting DC maturation. Use this validated flow cytometry panel to identify and phenotype CD4+ T regulatory cells.