Regulatory T Cells

Correlation and regression analysis

Correlation and regression analysis

Correlation and regression of responses, CD4+CD25(int/high)FoxP3+ vs. CD4+CD25(int/high)CD127low predictors in a PBMC assay.

Enrichment of Tregs with CD4, CD25, and CD127

Enrichment of Tregs with CD4, CD25, and CD127

PBMCs were stained with anti-human CD4, CD25, and CD127 as well as FoxP3. Lymphocytes were determined using the light scatter properties (FSC and SSC) and then gated on CD4 vs. CD25 as shown. Percentages of Tregs (FoxP3+) are shown for each of the CD25 populations, CD25high, CD25int, CD25low, and CD25-.

The BD Pharmingen brand Human Regulatory T Cell Cocktail

The BD Pharmingen brand Human Regulatory T Cell Cocktail

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 BD Pharmingen™ Human FoxP3 Buffer Set (Cat. No. 560098) and stained with BD Horizon™ V450 conjugated anti-human FoxP3 monoclonal antibody (Cat. No. 560459). 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 CD25high/CD127low T regulatory cells (solid line) and other T cells (dashed line). Flow cytometry was performed on a BD™ LSR II system.

Profile of anti-CD152 (CTLA-4) staining on fixed PBMCs

Profile of anti-CD152 (CTLA-4) staining on fixed PBMCs

PBMCs were surface stained with anti-CD4 PerCP Cy™5.5 (clone SK3, Cat. No. 341654) and anti-CD25 PE (clone 2A3, Cat. No. 341010). Cells were washed with staining buffer and fixed with BD Cytofix™ buffer (Cat. No. 554655 ) per the protocol. The cells were then permeabilized with BD Perm/Wash™ buffer (Cat. No. 554723), stained with anti-CD152 APC (clone BNI3, Cat. No. 555855), and acquired on a BD FACSCalibur™ system.

A) shows the gated events based on light scattering profile of lymphocytes and fluorescence characteristics of CD4 and CD25. The cells are differentiated as CD25high, CD25int,and CD25- based on their CD25 expression.

B) shows the overlay of the CD152 expression on these three subsets.