BD Pharmingen™ Human Regulatory T Cell Sorting Kit

The immune system protects individuals from a broad range of pathogenic microorganisms while avoiding inappropriate or extreme immune responses, such as autoimmune or allergic immune responses that could be harmful. Adaptive immunity is largely mediated by T and B lymphocyte populations that can specifically recognize a diverse constellation of foreign antigens. Through processes of rapid clonal expansion and differentiation, antigen-stimulated lymphocytes generate a vast array of potent effector and memory cell progeny to successfully rid the body of the offending foreign antigen. A distinct class of T cells, called regulatory T cells (Treg), safeguards against the immune system's responsiveness to self antigens and restrains excessive responsiveness to foreign antigens that could be harmful to the host.  Natural CD4+ T regulatory cells (nTreg) are generated in the thymus as a functionally mature subpopulation of T cells. These cells emigrate to the peripheral T cell pool and constitutively express medium to high levels of CD25 (IL-2 receptor α chain) and Foxp3 (forkhead box P3). The transcription factor Foxp3 is a member of the forkhead/winged-helix family of transcription factors and is a master regulator of Treg development and function. CD127 (IL-7 receptor α chain) constitutes another useful cell surface marker for identifying Treg cells. CD127 is downregulated on Treg cells, inversely correlating with the expression of Foxp3. Inducible CD4+ T regulatory cells (iTreg) can arise from mature peripheral naïve CD4+ T cells not expressing CD25 and Foxp3 (CD4+CD25-Foxp3- cells). Recent studies indicate that natural human CD4+FoxP3+ T cells can be subdivided into phenotypically- and functionally-distinct subsets based on the coexpressed levels of Foxp3, CD25 and CD45RA. The CD45RA marker is particularly useful for identifying resting CD4+ natural Treg cells (CD45RA+) as well as in vivo-activated CD4+ Treg (CD45RA-) when used with Foxp3, CD25, and CD127 markers. Phenotypic and functional studies using these markers provide a basis for high resolution analyses of the dynamics of Treg cell generation and differentiation in development, age and disease states. The BD™ Human Regulatory T Cell Sorting Kit provides reagents that allow investigators to stain viable human peripheral blood mononuclear cells (PBMC) or an enriched CD4+ T cell fraction of PBMC and to purify CD4+CD25+CD127lo cells (optional CD45RA+). These cells can then be tested functionally or used as precursors for expansion and differentiation cultures to generate activated Treg cells. Other cellular fractions such as CD4+CD25-CD127+ cells (naïve CD4+ T cells) can simultaneously be purified by flow cytometric sorting for subsequent functional characterization. In addition, fluorescent anti-human CD45RA antibody is included to facilitate further characterization or sorting of naïve (CD45RA+) or activated/memory (CD45RA-) CD4+Foxp3+ or CD4+Foxp3- T lymphocytes.

(RUO)

Flow cytometry, Fluorescence activated cell sorting (Tested During Development)

CD45RA, CD127, CD25, CD4

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