BD Biosciences Expands Regulatory T Cell Offerings
As part of its ongoing efforts to support research in T cell regulation, BD Biosciences has expanded its portfolio with three new high quality research reagents. TGF-β1, CD39 (Clone TÜ66), and a new cocktail.
Human TGF-β1 monoclonal antibody: a powerful new tool for studying adaptive Treg populations.
BD Pharmingen™-brand anti-human TGF-β1 PE conjugates are optimized for flow cytometric detection of TGF-β1, a member of the TGF-β superfamily. TGF-β1 is a key cytokine involved in a multitude of cellular processes and has recently been identified as playing an important role in the development and function of Regulatory T cells (Tregs).
| Description | Size | Cat. No. |
|---|---|---|
| BD Pharmingen PE Mouse anti-Human TGF-β (Clone: TB21; Mouse IgG1, κ) | 100 tests | 560227 |
Figure 1. TGF-β expression in activated human PBMCs.
Human PBMCs were activated with PMA and ionomycin, in the presence of Monensin and Brefeldin A (BFA) for 24 hours at 37°C. Activated cells were processed by fixation and permeabilization followed by staining with FITC-conjugated IFN-γ (Cat. No. 554551) and PE-conjugated TGF-β (Cat. No. 560227) monoclonal antibodies.
Human CD39 (clone TÜ66): enhanced characterization of Tregs
The BD Pharmingen-brand anti-human CD39 (clone TÜ66) monoclonal antibody is a recently identified marker for human Tregs and is available in PE and APC formats for use in flow cytometry. TÜ66 recognizes ENTPD1, an ectoenzyme that belongs to the family of ectonucleoside triphosphate diphosphohydrolases (E-NTPDases). Recently, it has been shown that CD39 is also expressed in FoxP3 expressing CD4+ Tregs in both human and mice, thereby adding to the list of cell surface markers such as CD25 and CD127 that could be used to identify CD4+ Tregs.1,2
| Description | Size | Cat. No. |
|---|---|---|
| BD Pharmingen PE Mouse Anti-Human CD39 (Clone: TÜ66; Mouse IgG2b, κ) | 100 tests | 555464 |
| BD Pharmingen APC Mouse Anti-Human CD39 (Clone: TÜ66; Mouse IgG2b, κ) | 100 tests | 560239 |
Figure 2. Analysis of human CD39 expression in whole blood
Whole blood samples were stained at room temperature in the dark for 20 minutes with FITC CD4 clone RPA-T4 (Cat. No. 555346), PE-Cy7 CD25 clone M-A251 (Cat. No. 557741), PE CD127 clone hIL-7R-M21 (Cat. No. 557938), and APC CD39 clone TÜ66 (Cat. No. 560239). The cells were then lysed and washed twice in 1% FBS wash buffer, and samples were run on a BD FACSCanto™ system.
References
1. Borsellino G, Kleinewietfeld M, Mitri D et al. Expression of ectonucleotidase CD39 by FOXP3+ Treg cells: hydrolysis of extracellular immune suppression. Blood. 2007;110:(4):1225-1232.
2. Deaglio S, Dwyer, KM, Gao W, et al. Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory T cells mediates immune suppression. J Exp Med. 2007;204:(6):1257-1265.
BD Human Regulatory T Cell Cocktail (CD4/CD25/CD127): simplify identification and isolation of Tregs
The BD Pharmingen-brand Human Regulatory T Cell Cocktail is a three-color reagent that can be used to identify CD4+CD25int/highCD127low live natural Treg populations. The reagent cocktail includes anti-human CD4 FITC (clone SK3), anti-human CD25 PE-Cy7 (clone 2A3), and anti-human CD127 Alexa Fluor® 647 (clone hIL-7R-M21). The one-step pre- mixed cocktail simplifies Treg identification and significantly enhances enrichment of live Treg populations by 2 to 4 times compared to gating on CD25high alone.
| Description | Clone | Isotype | Size | Cat. No. |
|---|---|---|---|---|
| BD Pharmingen™ Human Regulatory T cell cocktail | 100 tests | 560249 | ||
| CD4 FITC | SK3 | Mouse IgG1, κ | ||
| CD25 PE-Cy7 | 2A3 | Mouse IgG1, κ | ||
| CD127 Alexa Fluor® 647 | hIL-7R-M21 | Mouse IgG1, κ | ||
Figure 3. Three-color analysis of the expression of CD4, CD25, and CD127 on peripheral blood mononuclear cells (PBMCs).
PBMCs were stained with either an Isotype Control (Cat. No. 557872/555909, data not shown) or Human Regulatory T Cell Cocktail (Cat. No. 560249). The PBMCs were then fixed, lysed, and permeabilized using the BD Pharmingen Human FoxP3 Buffer Set (Cat. No. 560098) and stained with PE conjugated anti-human FoxP3 monoclonal antibody (Cat. No.560082). During data analysis, lymphocytes were identified by light scatter profile and CD4 positive expression. A) Data representing the CD25 and CD127 expression profile of the CD4 positive cells. B) Data showing hFoxP3 expression on CD127low/CD25high Tregs (solid line) and other T cells (dashed line). Flow cytometry was performed on a BD FACSCanto™ system.