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: 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

Journal: eLife

Article Title: SLAMF6​ deficiency augments tumor killing and skews toward an effector phenotype revealing it as a novel T cell checkpoint

doi: 10.7554/eLife.52539

Figure Lengend Snippet: ( A ) SLAMF6 and Vβ13 expression in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes measured by flow cytometry. ( B ) Percent CD8+, CD4, and CD19 cells in spleens from Pmel-1 or Pmel-1 x SLAMF6 -/- untreated mice. ( C ) Pmel-1, and Pmel-1 x SLAMF6 -/- CD8+ untreated splenocytes were stained with anti-CD44 and anti-CD62L. One representative experiment is shown. ( D ) Percent CD8+ cells in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes after 7 days of in vitro activation with gp100 25-33 peptide and IL-2 (30 IU/ml). ( E ) Flow cytometry for activation markers (CD25, CD69, CD137) in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes after 3 days of in vitro activation, as in ( D ). Median fluorescence intensity (MFI) is shown. ( F ) Expression of PD-1 in Pmel-1 or Pmel-1 x SLAMF6 -/- CD8+ T cells after 7 days of in vitro activation, as in ( D ). Median fluorescence intensity (MFI) is shown. ( G, H ) After 7 days of activation, Pmel-1 and Pmel-1 x SLAMF6 -/- CD8+ T cells were stained with anti-CD44 and anti-CD62L. CD8+ subpopulations were defined for each mouse strain. ( G ) One representative experiment and ( H ) summary of subpopulations identified by flow cytometry in five experiments is shown. EM, effector memory, CM, central memory. Student t-test. *, p<0.05, **, p<0.01, ***, p<0.001.

Article Snippet: Antibody , Monoclonal rat anti mouse CD25 (PC61.5) , Biogems, Westlake Village, CA , 07312 , 0.2 μg/100 μl.

Techniques: Expressing, Flow Cytometry, Staining, In Vitro, Activation Assay, Fluorescence

Journal: eLife

Article Title: SLAMF6​ deficiency augments tumor killing and skews toward an effector phenotype revealing it as a novel T cell checkpoint

doi: 10.7554/eLife.52539

Figure Lengend Snippet:

Article Snippet: Antibody , Monoclonal rat anti mouse CD25 (PC61.5) , Biogems, Westlake Village, CA , 07312 , 0.2 μg/100 μl.

Techniques: Generated, Immunohistochemistry, Sequencing, Recombinant, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Transformation Assay, Clonogenic Cell Survival Assay, Cell Isolation, Isolation, Software

Journal:

Article Title: Mechanism of Reduced T-Cell Effector Functions and Class-Switched Antibody Responses to Herpes Simplex Virus Type 2 in the Absence of B7 Costimulation

doi: 10.1128/JVI.77.4.2426-2435.2003

Figure Lengend Snippet: CD25 expression on CD4+ T cells. At 4 days following infection, (A) spleen and (B) genital lymph node cells from infected BALB/c and B7KO mice were analyzed by flow cytometry with antibodies specific for CD4 and CD25. Numbers within each quadrant represent the percentages of the cells within the lymphocyte gate. Cells from two to five mice were pooled. Data shown are from one of two experiments performed with similar results.

Article Snippet: Single-cell suspensions were prepared as described above, and cells (10 6 ) were stained with anti-CD4-fluorescein isothiocyanate or anti-CD8-fluorescein isothiocyanate (Caltag) and 0.5 μg of anti-CD25-phycoerythrin (Cedarlane, Ontario, Canada) or 1 μg of biotinylated anti-CD40L (PharMingen) followed by streptavidin-CyChrome in phosphate-buffered saline containing 0.2% bovine serum albumin and 0.1% azide.

Techniques: Expressing, Infection, Flow Cytometry