Description
Background information
Regulatory CD4+CD25+ T cells are suppressor cells that neutralize other immune cells by various mechanisms. Their characteristic marker is the transcription factor FoxP3. Regulatory CD4+CD25+ T cells seem to suppress harmful immunological reactions to self or foreign antigens.
Detailed separation procedure
The isolation is performed in a two-step procedure. First, non-CD4+ T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies against CD8, CD14, CD15, CD16, CD19, CD36, CD56, CD123, TCRγ/δ and CD235a (glycophorin A) and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS® Column. In the second step, the flow-through fraction of pre-enriched CD4+ T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+ regulatory T cells.
Downstream applications
The CD4+CD25+ Regulatory T Cell Isolation Kit has been used to investigate the role of CD4+CD25+ regulatory T cells in autoimmune diseases1,2, antitumor immunity3–6, allergy7, and in infectious diseases8. For further characterization, CD4+CD25+ regulatory T cells were isolated from patients suffering from rheumatoid arthritis1 or multiple sclerosis2, from acute T cell lymphoma patients4, from tumor biopsy specimens of non-Hodgkin lymphoma patients5, after dendritic cell vaccination9 as well as from patients with atopic dermatitis7.
Columns
For the first magnetic separation (depletion): LD or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
Overview
The CD4+CD25+ Regulatory T Cell Isolation Kit was developed for the isolation of CD4+CD25+ cells from human PBMCs. The isolation is performed in a two-step procedure.
