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.

Data and images for
CD4
+
CD25
+
Regulatory T Cell Isolation Kit
, human

Figures

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
PBMCs before separation
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
Pre-enriched CD4+ cells
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
Isolated CD4+CD25+ regulatory T cells
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
View details

Figure 1

CD4
+
CD25
+
regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.

Specifications for
CD4
+
CD25
+
Regulatory T Cell Isolation Kit
, human

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.

Detailed product information

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 diseases
1,2
, antitumor immunity
3–6
, allergy
7
, and in infectious diseases
8
. For further characterization, CD4
+
CD25
+
regulatory T cells were isolated from patients suffering from rheumatoid arthritis
1
or multiple sclerosis
2
, from acute T cell lymphoma patients
4
, from tumor biopsy specimens of non-Hodgkin lymphoma patients
5
, after dendritic cell vaccination
9
as well as from patients with atopic dermatitis
7
.

Columns

For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

References for
CD4
+
CD25
+
Regulatory T Cell Isolation Kit
, human

Publications

  1. Ehrenstein, M. R. et al. (2004) Compromised function of regulatory T cells in rheumatoid arthritis and reversal by anti-TNFalpha therapy. J. Exp. Med. 200: 277-285
  2. Hong, J. et al. (2006) CD4+ regulatory T cell responses induced by T cell vaccination in patients with multiple sclerosis. Proc. Natl. Acad. Sci. U.S.A. 103: 5024-5029
  3. Nishikawa, H. et al. (2006)
    Influence of CD4
    +
    CD25
    +
    regulatory T cells on low/high-avidity CD4
    +
    T cells following peptide vaccination.
    J. Immunol. 176: 6340-6346
  4. Karube et al. (2004) Expression of FoxP3, a key molecule in CD4CD25 regulatory T cells, in adult T-cell leukaemia/lymphoma cells. Br J Haematol 126: 81-84
  5. Yang et al. (2006)
    Intratumoral CD4
    +
    CD25
    +
    regulatory T-cell-mediated suppression of infiltrating CD4
    +
    T cells in B-cell non-Hodgkin lymphoma.
    Blood 107: 3639-3646
  6. Mougiakakos, D. et al. (2009) Naturally occurring regulatory T cells show reduced sensitivity toward oxidative stress-induced cell death. Blood 113: 3542-3545
  7. Ou, L. S. et al. (2004) T regulatory cells in atopic dermatitis and subversion of their activity by superantigens. J. Allergy Clin. Immunol. 113: 756-763
  8. Xu, D. et al. (2006)
    Circulating and liver resident CD4
    +
    CD25
    +
    regulatory T cells actively influence the antiviral immune response and disease progression in patients with hepatitis B.
    J. Immunol. 177: 739-747

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CD4
+
CD25
+
Regulatory T Cell Isolation Kit
, human

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