cd25 pe mab  (Proteintech)

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4 + CD25 + Regulatory T Cells

doi: 10.1155/2012/647308

Figure Lengend Snippet: Schematic diagram of the experimental protocol. On days 0 to 14, mice were divided into control (PBS-treated) and OVA-induced asthmatic groups. On day 18, OVA-induced asthmatic group was separated into 4 groups (OVA-immunized group; EA-treated after OVA induction group; anti-CD25 antibody injection after OVA induction group; EA-treated after OVA induction; anti-CD25 antibody injection group). Mice were immunized i.p. with 0.25 mg of anti-CD25 antibody 3 days before intranasal challenge. On day 21, all of OVA-induced asthmatic groups received 1% OVA in 50 μ L PBS or PBS intranasally and after 10 minutes, EA treatments at ST36 were applied for another 10 minutes. Acupuncture stimulations were repeated for ten days consecutively after intranasal challenge.

Article Snippet: Single-cell suspensions and pneumonocytes obtained from Foxp3EGFP Balc/C mice were labeled with anti-CD4-APC and anti-CD25-PE mAb using standard staining methods, and the percentage of cells stained with a particular reagent was analyzed by FACSCalibur using the CellQuest software (BD Bioscience).

Techniques: Injection

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4 + CD25 + Regulatory T Cells

doi: 10.1155/2012/647308

Figure Lengend Snippet: Confirmation of CD4 + CD25 + Treg cells depletion. PBS treatment alone; anti-CD25 antibody injection into WT Balb/c Foxp3 + EGFP (anti-CD25). Balb/c mice received injections of 0.25 mg of anti-CD25 antibody after OVA sensitization. The efficacy of CD4 + CD25 + Treg cells depletion was confirmed by flow cytometry analysis using, PE-anti-mouse CD25 and Foxp3 + EGFP . Data are shown as mean ± S.E.M. (*** P < 0.001 versus control n = 4).

Article Snippet: Single-cell suspensions and pneumonocytes obtained from Foxp3EGFP Balc/C mice were labeled with anti-CD4-APC and anti-CD25-PE mAb using standard staining methods, and the percentage of cells stained with a particular reagent was analyzed by FACSCalibur using the CellQuest software (BD Bioscience).

Techniques: Injection, Flow Cytometry

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4 + CD25 + Regulatory T Cells

doi: 10.1155/2012/647308

Figure Lengend Snippet: Flow cytometry analysis of splenic CD4 + CD25 + Foxp3 + Treg cells. WT Foxp3 EGFP mice were sacrificed after 10 days of EA stimulation. Isolated splenocytes were stained with anti-CD4 allophycocyanin (APC) and anti-CD25 PE and submitted to flow cytometry analysis. At first, CD4 + cells were gated (not shown), and then CD25 + and Foxp3 EGFP-positive cells were analyzed. The average flow cytometry analysis data of CD4 + CD25 + Foxp3 + Treg cells (WT 4.28%; EA-treated at ST36 8.25%; EA-treated at tail 6.79%) were represented. Data are shown as mean ± S.E.M. (* P < 0.05; ** P < 0.01 versus WT; n = 5).

Article Snippet: Single-cell suspensions and pneumonocytes obtained from Foxp3EGFP Balc/C mice were labeled with anti-CD4-APC and anti-CD25-PE mAb using standard staining methods, and the percentage of cells stained with a particular reagent was analyzed by FACSCalibur using the CellQuest software (BD Bioscience).

Techniques: Flow Cytometry, Isolation, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Attenuates Ovalbumin-Induced Allergic Asthma via Modulating CD4 + CD25 + Regulatory T Cells

doi: 10.1155/2012/647308

Figure Lengend Snippet: Effect of EA on CD4 + CD25 + Foxp3 EGFP Treg cells and Treg cells depletion in OVA-induced asthmatic pneumonocytes. PBS treatment alone (control); OVA induction only (OVA); EA treatment after OVA induction (OVA + EA); anti-CD25 Ab injection after OVA induction (OVA_T); EA stimulation after OVA induction and anti-CD25 Ab injection (OVA_T + EA). Isolated pneumonocytes were stained with anti-CD4 allophycocyanin and anti-CD25 PE and submitted to flow cytometry analysis. CD4 + cells were gated on and then CD25 + Foxp3 EGFP-positive cells were examined. The average flow cytometry analysis data of all groups were depicted. There was an increase of CD4 + CD25 + Foxp3 + Treg cells in OVA+EA group, but negligible data were shown in anti-CD25 Ab injected groups when compared with OVA group. Data are shown as mean ± S.E.M. (*** P < 0.001 versus control; ** P < 0.01 versus OVA; n = 6).

Article Snippet: Single-cell suspensions and pneumonocytes obtained from Foxp3EGFP Balc/C mice were labeled with anti-CD4-APC and anti-CD25-PE mAb using standard staining methods, and the percentage of cells stained with a particular reagent was analyzed by FACSCalibur using the CellQuest software (BD Bioscience).

Techniques: Injection, Isolation, Staining, Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: TCR-triggered extracellular superoxide production is not required for T-cell activation

doi: 10.1186/s12964-014-0050-1

Figure Lengend Snippet: Extracellular superoxide production is not required for primary human T-cell activation. (A) Primary human T cells were stimulated with CD3xCD28-coated microbeads alone or in the presence of either SOD, catalase or ascorbate. After 16 hours cells were stained with CD25-FITC and CD69-PE mAbs and analyzed by flow cytometry. The data are representative of 3 independent experiments. (B) Quantification of (A) . The values indicate the mean fluorescence intensities normalized to stimulated controls. (C) Quantification of (A) . The values indicate the percentages of CD25 + CD69 + cells normalized to stimulated controls.

Article Snippet: After 16 h, cells were stained with FITC- or PE-labeled mAbs against CD25 and CD69 (BD Pharmingen, BioLegend) and analyzed by flow cytometry using a FACSCalibur and CellQuest software or BD LSRFortessa and FACSDiva Software 6.1.3 (all from BD Biosciences), and FlowJo 7.5.5 (Tree Star, Inc.).

Techniques: Activation Assay, Staining, Flow Cytometry, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: TCR-triggered extracellular superoxide production is not required for T-cell activation

doi: 10.1186/s12964-014-0050-1

Figure Lengend Snippet: NOX2 is not required for primary T-cell activation. (A) Splenic T cells from WT or gp91 phox−/− mice were stimulated with CD3xCD28-coated microbeads. After 16 hours cells were stained with CD25-FITC and CD69-PE mAbs and analyzed by flow cytometry. The data are representative of at least 2 independent experiments. 1 WT and 2 gp91 phox−/− mice were used in each experiment. (B) Quantification of (A) . The values indicate the mean fluorescence intensities. (C) Quantification of (A) . The values indicate the percentages of CD69 + CD25 + cells.

Article Snippet: After 16 h, cells were stained with FITC- or PE-labeled mAbs against CD25 and CD69 (BD Pharmingen, BioLegend) and analyzed by flow cytometry using a FACSCalibur and CellQuest software or BD LSRFortessa and FACSDiva Software 6.1.3 (all from BD Biosciences), and FlowJo 7.5.5 (Tree Star, Inc.).

Techniques: Activation Assay, Staining, Flow Cytometry, Fluorescence