Journal: Scientific Reports

Article Title: Tuning IL-2 signaling by ADP-ribosylation of CD25

doi: 10.1038/srep08959

Figure Lengend Snippet: Splenocytes from DEREG mice were incubated in the absence or presence of the indicated concentrations of NAD + before staining with fluorochrome-conjugated mAbs directed against CD4 and CD25 (mAb PC61 or mAb 7D4). Gating was performed on CD4 + cells. (a) Representative dot plots of cells incubated in the absence or presence of 12 μM NAD + . (b) Percentages of CD4 + GFP + cells staining with anti-CD25 mAbs plotted as a function of the concentration of added NAD + . Results are representative of two independent experiments.

Article Snippet: Alternatively, after erythrocyte lysis with Ack lysis buffer (Bio Whittaker) and depletion of B-cells with Dynabead-conjugated sheep anti-mouse IgG (Invitrogen), Tregs were positively selected by magnetic cell sorting using PE conjugated anti-CD25 (7D4) and magnetic bead-conjugated anti-PE antibodies according to the manufacturer's instructions (Miltenyi Biotec).

Techniques: Incubation, Staining, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Establishing Porcine Monocyte-Derived Macrophage and Dendritic Cell Systems for Studying the Interaction with PRRSV-1

doi: 10.3389/fmicb.2016.00832

Figure Lengend Snippet: Table of antibodies.

Article Snippet: CD25 , Mouse , Pig , K231.3B2 , IgG1 , Unconjugated , AbD Serotec.

Techniques:

Journal: Journal of Inflammation Research

Article Title: Cerium Oxide-Loaded Exosomes Derived From Regulatory T Cells Ameliorate Inflammatory Bowel Disease by Scavenging Reactive Oxygen Species and Modulating the Inflammatory Response

doi: 10.2147/JIR.S502388

Figure Lengend Snippet: Synthesis and characterization of Treg-exo@nCeO. ( A ) TEM images of nCeO, scale bars: 10 nm. ( B ) EDS spectrum of nCeO. ( C ) Appearance images of nCeO. ( D – F ) hydrogen peroxide and free radical scavenging assay, n=3; *p < 0.05 vs 0 μg/mL, **p < 0.01 vs 0 μg/mL, ns, no significance vs 0 μg/mL. ( G ) Flow chart of the isolation of CD4+CD25+ T cells. ( H ) Relative proportion of living cells, n=3; **p < 0.01 vs 0 μg/mL, ns, no significance vs 0 μg/mL. ( I ) Uptaken Cy5-labeled nCeO and fluorescence imaging, scale bars: 2 μm. ( J ) NTA of exo and exo@nCeO. ( K ) Western blot assay of the expression of Calnexin, Tsg101 and CD63 in Tregs and exo. ( L ) TEM images and average size of exo and exo@nCeO, n=3, scale bars: 100 nm. ( M ) Uptaken Dio-exo@Cy5-nCeO and fluorescence imaging, scale bars: 5 μm.

Article Snippet: 1×10 6 Tregs were collected and stained with phycoerythrin (PE) monoclonal antibodies against mouse CD25 (E-AB-F1102D, Elabscience, China) and fluorescein isothiocyanate (FITC) conjugated monoclonal antibodies against mouse CD4 (E-AB-F1097C, Elabscience, China) for 30 min, followed by data collection on a flow cytometer (Beckman Coulter, USA).

Techniques: Isolation, Labeling, Fluorescence, Imaging, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Fucoxanthin Ameliorates Atopic Dermatitis Symptoms by Regulating Keratinocytes and Regulatory Innate Lymphoid Cells

doi: 10.3390/ijms21062180

Figure Lengend Snippet: The characterization of Il-2 producers regulated by FX and TAC. FX stimulated Il-2 production in CD45R + CD25 + ILCregs. Some Il-2 + CD45R + CD25 + ILCregs were observed in TAC treated skin and others were CD45R - CD25 - cells. The panel is focused on the dermis.

Article Snippet: Sections were subjected to immunohistochemistry using antibodies of Alexa Fluor 488 ® -anti-mouse Il-10 (#505013), Brilliant Violet 421TM-anti-mouse TGF-β1 (#141407), Pacific BlueTM-anti-mouse IL-2 (#503820), Alexa Fluor ® 488-anti-GATA3 (#653808), PE-anti-mouse Ly-6A/E (Sca-1) (#108108), PE-anti-mouse IL-33Rα (ST2) (#146608) and PE-anti-mouse/human IL-5 (#504304) (BioLegend, San Diego, CA, USA), FITC-anti-mouse/human CD45R (#11-0452-82) (Thermo Fisher Scientific, Waltham, MA, USA), PE-anti-mouse CD25 (#130-120-766) (Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques:

Journal:

Article Title: Herpes Simplex Virus Type 1-Specific Cytotoxic T-Lymphocyte Arming Occurs within Lymph Nodes Draining the Site of Cutaneous Infection

doi:

Figure Lengend Snippet: Vβ10 expression by CD8+ CD25+ activated T cells from the draining lymph nodes of HSV-1-infected mice determined directly ex vivo or after 3 days of culture without exogenous antigen. Three mice were infected in the hind footpads with 4 × 105 PFU of HSV-1, and 5 days later the cells from the draining popliteal lymph nodes were collected, viable cell counts were performed, and 1 × 106 to 2 × 106 cells were placed into culture without exogenous antigen. Another 106 cells were triple stained with Vβ10, CD25, and CD8 antibodies prior to analysis by flow cytometry. After 3 days the cultured cells were similarly analyzed by flow cytometry. Dead cells were excluded by using propidium iodide staining. The dot plots represent CD8 versus CD25 staining for the popliteal lymph node cells ex vivo, and after 3 days in culture without exogenous antigen (in vitro). The histogram shows the Vβ10 receptor expression of the CD8+ CD25+ T cell subsets. The percentage of CD8+ CD25+ cells in the lymph nodes, and the percentage of Vβ10+ T cells among the CD8+ CD25+ T cell subsets are shown as a mean and standard deviation for the three mice analyzed.

Article Snippet: Lymph node cells cultured for 3 days without exogenous antigen were also stained with allophycocyanin-labeled anti-CD8 (53-6.7; Pharmingen) and FITC-labeled anti-CD25 (PC61.5.3; Cedarlane Laboratories, Ltd., Hornby, Ontario, Canada).

Techniques: Expressing, Infection, Ex Vivo, Staining, Flow Cytometry, Cell Culture, In Vitro, Standard Deviation

Journal:

Article Title: Herpes Simplex Virus Type 1-Specific Cytotoxic T-Lymphocyte Arming Occurs within Lymph Nodes Draining the Site of Cutaneous Infection

doi:

Figure Lengend Snippet: The CD8+ CD25+ T cells from the draining lymph nodes of HSV-1-infected mice analyzed after 3 days of culture without exogenous antigen are predominantly gB specific. A mouse was infected in the hind footpads with 4 × 105 PFU of HSV-1, and 5 days later the cells from the draining popliteal lymph nodes were collected and placed into culture without exogenous antigen. After 3 days, the cultured cells and lymph node cells isolated from a naive mouse were triple stained with CD25 and CD8 antibodies, and the H-2Kb/gB tetramer and analyzed by flow cytometry. Dead cells were excluded by using propidium iodide staining. The dot plot represents CD8 versus CD25 staining for the cultured popliteal lymph node cells. The histograms show the level of H-2Kb/gB tetramer staining of the CD8+ CD25hi and CD8+ CD25lo T cell subsets from the cultured (shaded) and naive (unshaded) lymph node cell populations.

Article Snippet: Lymph node cells cultured for 3 days without exogenous antigen were also stained with allophycocyanin-labeled anti-CD8 (53-6.7; Pharmingen) and FITC-labeled anti-CD25 (PC61.5.3; Cedarlane Laboratories, Ltd., Hornby, Ontario, Canada).

Techniques: Infection, Cell Culture, Isolation, Staining, Flow Cytometry

Journal: BMC Complementary Medicine and Therapies

Article Title: Effect and mechanisms of shikonin on breast cancer cells in vitro and in vivo

doi: 10.1186/s12906-024-04671-3

Figure Lengend Snippet: Shikonin regulates immunity in vivo. ( A and B ) Representative scatter plots of spleen CD25 + Foxp3 + positive T cells ( A ) and CD8 + positive T cells ( B ). ( C and D ) Statistical analysis of percentage of CD25 + Foxp3 + positive T cells ( C ) and CD4 + positive T cells ( D ). ( E ) Statistical analysis of CD8 + positive T cells percentage. ( F ) Statistical analysis of the ratio of CD4 + and CD8 + positive T cells. Data are presented as mean ± SEM ( n = 6); * P < 0.05, ** P < 0.01

Article Snippet: The other portion was incubated with Elab Fluor ® Violet 450 Anti-Mouse CD45 Antibody[30-F11], Elab Fluor ® Red 780 Anti-Mouse CD3 Antibody[17A2], APC Anti-Mouse CD4 Antibody[GK1.5], and PerCP/Cyanine5.5 Anti-Mouse CD25 Antibody[PC-61.5.3] (Elabscience, China) diluted in cell staining buffer at 4 °C in the dark for 30 min.

Techniques: In Vivo

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 2 | JHU083 inhibited T cells activation in vivo. Mice of Vehicle-AIH and JHU083-AIH group were intravenously injected with 8ml/kg ConA, and Vehicle-WT group was injected with normal saline of equal volume. JHU083-AIH group was gavaged with 0.3mg/kg JHU083 24h and 1h before ConA injection, and mice of Vehicle-WT and Vehicle-AIH group were treated with equal vehicle at the same time points. (A, B) Flow cytometry was applied to analyze the expression of CD25 and CD69 of CD4(+) (A) and CD8(+) (B) T cells. Data were expressed as means ± SEM. *P < 0.05 and **P <0.01.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: Activation Assay, In Vivo, Injection, Saline, Flow Cytometry, Expressing

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 4 | DON treatment suppressed T cells activation in vitro. Freshly separated spleen cells were stimulated with 1.5mg/ml ConA with or without treating 2.5mM DON, and cells were analyzed 24h after ConA stimulation. (A, B) The expressions of activation markers CD25 and CD69 in CD4(+) (A) and CD8(+) (B) T cells were examined by flow cytometry. Data were expressed as means ± SEM. **P <0.01, and ***P <0.001.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: Activation Assay, In Vitro, Cytometry

Journal: Frontiers in immunology

Article Title: Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.

doi: 10.3389/fimmu.2022.880262

Figure Lengend Snippet: FIGURE 7 | The function of DON to mTOR signaling may be mediated by amino transporter amino transporter SLC7A5. (A) qRT-PCR was applied to analyze amino acid transporters SLC7A5 and SLC1A5 mRNA levels. (B, C) Flow cytometry was applied to analyze the degree of activation (expression of CD25 and CD69) of CD4 (+) (B) and CD8(+) (C) T cells with or without treating LAT1-IN1. The levels of phosphorylated mTOR (D) as well as phosphorylated P70S6K (E) were examined by western blotting. Data were expressed as means ± SEM. *P < 0.05, **P <0.01, and ***P <0.001. ns, not significant.

Article Snippet: The antibodies used in the flow cytometry were as follows: CD4 (FITC, E-AB-F1097C), CD8a (PE, E-AB-F1104D), CD25 (PECy5, E-AB-F1102G), CD69 (PE-Cy7, E-AB-F1187H), IL4 (PE, E-AB-F1204UD), IL17A (APC, E-AB-F1272E) were purchased from Elabscience Biotechnology (Wuhan, China), antibodies IFNg (APC-Cy7, cat# 561479) was purchased from BD Biosciences (UK), and Granzyme B (PE-Cy7, cat# 372213) was purchased from Biolegend (San Diego, CA, USA).

Techniques: Quantitative RT-PCR, Flow Cytometry, Activation Assay, Expressing, Western Blot