pe cy7-labeled anti-cd25  (Thermo Fisher)

Journal: Frontiers in Immunology

Article Title: 20 S -Hydroxyvitamin D3, a Secosteroid Produced in Humans, Is Anti-Inflammatory and Inhibits Murine Autoimmune Arthritis

doi: 10.3389/fimmu.2021.678487

Figure Lengend Snippet: Treatment of CIA mice with 20 S (OH)D 3 reduces percentages of CD 4 + T cells and B cells in draining lymph nodes while elevating FoxP3 Treg/CD3 + CD4 + ratio*.

Article Snippet: Isolated cells from spleen or draining para aortic, popliteal or inguinal lymph nodes were labeled as follows: Alexa-700 labeled rat anti-mouse CD3, PE-Cy7-labeled rat anti-mouse CD25, Per CP-Cy5.5-labeled rat anti-mouse CD4, FoxP3 Tregs were detected by FoxP3 staining kit, and FITC-labeled rat anti-mouse CD-19 (BD Bioscience, San Diego, CA).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Self-specific memory regulatory T cells protect embryos at implantation in mice

doi: 10.4049/jimmunol.1202413

Figure Lengend Snippet: (A) Treg ablation was achieved by anti-CD25 mAb treatment administered one day prior to the mating of CBA/J mice. Histograms depict the % of fetal resorption in the indicated groups. The results are shown as normalized to the mean resorption frequency of the PBS-treated CBA x DBA control group, which was set at 100%. Numbers above bars indicate the cumulative numbers of resorbed/viable fetuses and the numbers of mice per experimental condition on dpi 8. Two-tailed Mann-Whitney test: * P < 0.05, ** P < 0.01. (B) Evaluation % of fetus resorption after fetal antigen preimmunization of the indicated mice with or without anti-CD25 mAb-induced Treg depletion at the time of the mating. BALB/cfemale first received anti-HA naïve CD4+CD25− Teffs from SFE donor mice (i.v.), and were then immunized by HA peptide (s.c., CFA condition). 2 months later, HA-immunized SFE-transferred BALB mice, similarly HA-immunized SFE mice and naïve SFE and BALB/c female were mated with pgkHA males directly or after Treg depletion by anti-CD25 treatment (- Treg condition). Viable fetuses or resorption sites were counted on dpi 8. (n = 2–4 mice per group). Two-tailed unpaired t-test: * P < 0.05, ** P < 0.01.

Article Snippet: Para-aortic and brachial LNs were mechanically dissociated in PBS (3% FBS) and stained with FITC- or V500-labeled anti-CD4, APC-labeled anti-CD8, PerCP-labeled anti-Thy1.1/CD90.1, biotin-labeled anti-CD62L (all from BD Biosciences), PE-Cy7-labeled anti-CD25, FITC-labeled anti-CD44, PE-labeled anti-PD-L1, APC-labeled anti-CD103 or PE-labeled anti-GITR, APC-labeled anti-CTLA-4 and PE-Cy5-labeled anti-ICOS mAbs (all from eBiosciences) The PE-labeled clone 6.5 anti-clonotypic mAb specific to TCR-HA was produced in rats immunized with the soluble TCR (a kind gift from Dr. H. von Boehmer, Dana-Farber Cancer Institute).

Techniques: Two Tailed Test, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Self-specific memory regulatory T cells protect embryos at implantation in mice

doi: 10.4049/jimmunol.1202413

Figure Lengend Snippet: (A) Effects of IL-2 treatment on Treg population of female CBA/J mice. Percentages of CD103 and Ki67 expressing Tregs and CD25 and PD-L1 mean fluorescence index (MFI) of Tregs in para-aortic and brachial LNs after a 10-day treatment with IL-2 (n = 5) or PBS (n = 5). (B-C) Effect of rhFlt3-L and IL-2 treatment on the outcome of CBA/J x DBA/2J matings.

Article Snippet: Para-aortic and brachial LNs were mechanically dissociated in PBS (3% FBS) and stained with FITC- or V500-labeled anti-CD4, APC-labeled anti-CD8, PerCP-labeled anti-Thy1.1/CD90.1, biotin-labeled anti-CD62L (all from BD Biosciences), PE-Cy7-labeled anti-CD25, FITC-labeled anti-CD44, PE-labeled anti-PD-L1, APC-labeled anti-CD103 or PE-labeled anti-GITR, APC-labeled anti-CTLA-4 and PE-Cy5-labeled anti-ICOS mAbs (all from eBiosciences) The PE-labeled clone 6.5 anti-clonotypic mAb specific to TCR-HA was produced in rats immunized with the soluble TCR (a kind gift from Dr. H. von Boehmer, Dana-Farber Cancer Institute).

Techniques: Expressing, Fluorescence

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

doi: 10.1590/1414-431X20143483

Figure Lengend Snippet: Multi-parameter strategy of analysis to identify TREG cells. A representative healthy control sample was analyzed by flow cytometry according to the expression of CD4, CD25, CD127, and FoxP3. CD25 + and CD25 high cell subsets presented distinct frequency of CD127 Ø/low cells and in both subsets the CD127 Ø/low phenotype was associated with high FoxP3 expression.

Article Snippet: PBMCs were washed in phosphate-buffered saline and 0.5×10 6 cells were incubated with fluorescein isothiocyanate (FITC) or allophycocyanin (APC)-labeled anti-CD127, APC-Cy3-labeled anti-CD3, peridinin chlorophyll (PerCP)-labeled anti-CD4, and phycoerythrin (PE)-Cy7-labeled anti-CD25 antibodies (Becton Dickinson, USA), according to the manufacturer's instructions.

Techniques: Flow Cytometry, Expressing

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

doi: 10.1590/1414-431X20143483

Figure Lengend Snippet: Analysis of the frequency of TREG-I (CD25 + CD127 Ø/low FoxP3 + ) and TREG-II (CD25 high CD127 Ø/low FoxP3 + ) cells expressing CD28 ( A ), CD40L ( B ), and CTLA-4 ( C ) in patients with active SLE (A-SLE), inactive SLE (I-SLE), and healthy controls (HC). The relative frequency of CD28 + , CD40L + , and CTLA-4 + cells was calculated over total TREG-I and TREG-II, respectively. Horizontal bars indicate means and standard deviation, respectively. *P<0.05; **P<0.01; ***P<0.001 (Kruskal-Wallis test).

Article Snippet: PBMCs were washed in phosphate-buffered saline and 0.5×10 6 cells were incubated with fluorescein isothiocyanate (FITC) or allophycocyanin (APC)-labeled anti-CD127, APC-Cy3-labeled anti-CD3, peridinin chlorophyll (PerCP)-labeled anti-CD4, and phycoerythrin (PE)-Cy7-labeled anti-CD25 antibodies (Becton Dickinson, USA), according to the manufacturer's instructions.

Techniques: Expressing, Standard Deviation

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

doi: 10.1590/1414-431X20143483

Figure Lengend Snippet: Correlation between the relative frequency of CD40L + TREG-I cells and disease activity (SLEDAI) in patients with SLE. The relative frequency of CD40L + TREG-I cells was calculated over total TREG-I (CD25 + CD127 Ø/low FoxP3 + ). The Spearman correlation test was used for analysis.

Article Snippet: PBMCs were washed in phosphate-buffered saline and 0.5×10 6 cells were incubated with fluorescein isothiocyanate (FITC) or allophycocyanin (APC)-labeled anti-CD127, APC-Cy3-labeled anti-CD3, peridinin chlorophyll (PerCP)-labeled anti-CD4, and phycoerythrin (PE)-Cy7-labeled anti-CD25 antibodies (Becton Dickinson, USA), according to the manufacturer's instructions.

Techniques: Activity Assay

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

doi: 10.1590/1414-431X20143483

Figure Lengend Snippet: Analysis of the frequency of TREG-I (CD25 + CD127 low/Ø FoxP3 + ) and TREG-II (CD25 high CD127 low/Ø FoxP3 + ) cells expressing OX40 ( A ), PD1 ( B ), and GITR ( C ) in patients with active SLE (A-SLE), inactive SLE (I-SLE), and healthy controls (HC). The relative frequency of OX40 + , PD1 + , and GITR + cells was calculated over total TREG-I and TREG-II, respectively. Horizontal bars indicate means and standard deviation, respectively. *P<0.05 (Kruskal-Wallis test).

Article Snippet: PBMCs were washed in phosphate-buffered saline and 0.5×10 6 cells were incubated with fluorescein isothiocyanate (FITC) or allophycocyanin (APC)-labeled anti-CD127, APC-Cy3-labeled anti-CD3, peridinin chlorophyll (PerCP)-labeled anti-CD4, and phycoerythrin (PE)-Cy7-labeled anti-CD25 antibodies (Becton Dickinson, USA), according to the manufacturer's instructions.

Techniques: Expressing, Standard Deviation

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Imbalanced expression of functional surface molecules in regulatory and effector T cells in systemic lupus erythematosus

doi: 10.1590/1414-431X20143483

Figure Lengend Snippet: Ratio of TREG-I (CD25 + CD127 Ø/low FoxP3 + ) or TREG-II (CD25 high CD127 Ø/low FoxP3 + ) cells to effector T cells expressing CD45RO ( A ), HLA-DR ( B ), GITR ( C ), OX40 ( D ), CD40L ( E ), and PD1 ( F ) in patients with active SLE (A-SLE), inactive SLE (I-SLE), and healthy controls (HC). Horizontal bars indicate means and standard deviation, respectively. Teff: effector T cells. *P<0.05 (Kruskall-Wallis test).

Article Snippet: PBMCs were washed in phosphate-buffered saline and 0.5×10 6 cells were incubated with fluorescein isothiocyanate (FITC) or allophycocyanin (APC)-labeled anti-CD127, APC-Cy3-labeled anti-CD3, peridinin chlorophyll (PerCP)-labeled anti-CD4, and phycoerythrin (PE)-Cy7-labeled anti-CD25 antibodies (Becton Dickinson, USA), according to the manufacturer's instructions.

Techniques: Expressing, Standard Deviation