Journal: iScience

Article Title: Defects in NK cell immunity of pediatric cancer patients revealed by deep immune profiling

doi: 10.1016/j.isci.2024.110837

Figure Lengend Snippet:

Article Snippet: Anti-CD25 (clone 2A3, conjugated to 149Sm) , Standard BioTools , Cat#3149010B.

Techniques: Purification, Clinical Proteomics, Recombinant, Blocking Assay, Staining, Saline, Mass Cytometry, Software, Cytometry

Journal: Human Vaccines & Immunotherapeutics

Article Title: Research and development of mAb drugs in China: a look from policy perspectives

doi: 10.1080/21645515.2019.1605809

Figure Lengend Snippet: Domestically approved mAb drugs by 2017.

Article Snippet: , Jiannipai , Recombinant Humanized Anti-CD25 mAbs Injection , Shanghai CP Guojian Pharmaceutical Co., Ltd. , Humanzied , 2011 , Transplant Rejection.

Techniques: Injection, Cream, Recombinant

Journal: iScience

Article Title: DUSP6 deletion protects mice and reduces disease severity in autoimmune arthritis

doi: 10.1016/j.isci.2024.110158

Figure Lengend Snippet:

Article Snippet: anti-CD25-APC (clone: 4E3) , Miltenyi Biotec, Germany , Cat# 130-113-846.

Techniques: Transgenic Assay, Knock-Out

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: International Journal of Molecular Sciences

Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2

doi: 10.3390/ijms20225744

Figure Lengend Snippet: Stimulation assay of chIL-2 monomer and dimer in vivo. ( A ) and ( B ), immunofluorescence analysis of CD4 + and CD25 + T cells by flow cytometry using FITC-conjugated anti-CD4 (CD4 FITC) and Alexa Fluor 647-conjugated anti-CD25 (CD25 AF647) antibodies before ( A ) or post ( B ) stimulation of chIL-2. ( C ), the bar graph of CD4 + T cell proportions; ( D ), the bar graph of CD25 + T cell proportions; BI, chIL-2 protein was not treated with incubation at 37 °C; PI, chIL-2 was treated with incubation at 37 °C. The plotted data represent mean values ± standard deviation, and are the average of three independent experiments.

Article Snippet: The isolated T cells from in vivo or in vitro experiments were then utilized to evaluate the proportion of CD4 + or CD25 + T cells among the totally counted cells or T cells (CD3 was used as T cell marker.) by flow cytometry with the following antibodies: PE-conjugated anti-chicken CD3 antibody, FITC-conjugated anti-chicken CD4 antibody (Southern Biotech, Birmingham, AL, USA), and Alexa Fluor 647 (AF647) conjugated anti-chicken CD25 antibody (Bio-Rad Labs, Inc. Hercules, CA, USA).

Techniques: In Vivo, Immunofluorescence, Flow Cytometry, Incubation, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2

doi: 10.3390/ijms20225744

Figure Lengend Snippet: Binding assays between chIL-2 monomer/dimer and chIL-2R in vivo. ( A ), ( B ) and ( C ) were immunofluorescence analyses of the T cells bound hexa-His tagged monomeric or dimeric chIL-2 by flow cytometry using FITC-conjugated anti-CD4 (CD4 FITC) and Alexa Fluor 647-conjugated anti-hexa-His (6×HisTag AF647) antibodies before ( A ) or post stimulation of monomeric ( B ) or dimeric ( C ) chIL-2. The proportions of detected T cells with each antibody were plotted as a bar graph ( D ). Ctrl, the proportions of T cells without the stimulation of any form of chIL-2; Monomer, the proportions of T cells that stimulated by sf9 source of monomeric chIL-2 protein; Dimer, the proportions of T cells that stimulated by sf9 source of dimeric chIL-2 protein. The plotted data represent mean values ± standard deviation, and are the average of three independent experiments.

Article Snippet: The isolated T cells from in vivo or in vitro experiments were then utilized to evaluate the proportion of CD4 + or CD25 + T cells among the totally counted cells or T cells (CD3 was used as T cell marker.) by flow cytometry with the following antibodies: PE-conjugated anti-chicken CD3 antibody, FITC-conjugated anti-chicken CD4 antibody (Southern Biotech, Birmingham, AL, USA), and Alexa Fluor 647 (AF647) conjugated anti-chicken CD25 antibody (Bio-Rad Labs, Inc. Hercules, CA, USA).

Techniques: Binding Assay, In Vivo, Immunofluorescence, Flow Cytometry, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Four Cysteine Residues Contribute to Homodimerization of Chicken Interleukin-2

doi: 10.3390/ijms20225744

Figure Lengend Snippet: Pull-down assay for characterization of the interaction between chIL-2 and CD25 in vivo. Hexa-His tagged chIL-2 was purified from the T cells that were isolated from hexa-His tagged chIL-2 treated chicken. Monomeric (lower panel in A and B ) or dimeric (lower panel in C and D ) chIL-2 were detected by anti-chIL-2 antibody, respectively. CD25 protein was pulled down along with monomeric or dimeric chIL-2 purification, and detected by anti-CD25 antibody (upper panel in A , B , C and D ). D-chIL-2 and M-chIL-2 are the recombinant chIL-2 monomer and dimer purified from Sf9, respectively. M1~M15, chIL-2 mutants with different Cys mutation combinations (refer to B and ). Marker, protein standard marker. WT, wild type chIL-2.

Article Snippet: The isolated T cells from in vivo or in vitro experiments were then utilized to evaluate the proportion of CD4 + or CD25 + T cells among the totally counted cells or T cells (CD3 was used as T cell marker.) by flow cytometry with the following antibodies: PE-conjugated anti-chicken CD3 antibody, FITC-conjugated anti-chicken CD4 antibody (Southern Biotech, Birmingham, AL, USA), and Alexa Fluor 647 (AF647) conjugated anti-chicken CD25 antibody (Bio-Rad Labs, Inc. Hercules, CA, USA).

Techniques: Pull Down Assay, In Vivo, Purification, Isolation, Recombinant, Mutagenesis, Marker

Journal: Arthritis Research & Therapy

Article Title: Therapeutic effect of a novel histone deacetylase 6 inhibitor, CKD-L, on collagen-induced arthritis in vivo and regulatory T cells in rheumatoid arthritis in vitro

doi: 10.1186/s13075-017-1357-2

Figure Lengend Snippet: Increased cytotoxic T-lymphocyte antigen-4 ( CTLA-4 ) expression by CKD-L in Treg cells from C57BL/6 mice ( n = 3). CD4 + CD25 – T cells were isolated from splenocytes of C57BL/6 mice. CD4 + CD25 – T cells were incubated with vehicle or HDAC6 inhibitors (1 to 10 μM) in the presence of antiCD3/CD28 beads and recombinant TGF-β2 in a 48-well plate for 6 days. CTLA-4 expression in CD4 + CD25 + Foxp3 + T cells was analyzed by mean fluorescence intensity ( MFI ; mean ± SEM) using flow cytometry. * p < 0.05, ** p < 0.001, vs vehicle

Article Snippet: Isolated CD4 + cells were incubated with microbead conjugated to a monoclonal antihuman CD25 antibody (MACS®; Miltenyi Biotec) for 15 min at 4 °C.

Techniques: Expressing, Isolation, Incubation, Recombinant, Fluorescence, Flow Cytometry

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: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD25 , REA945 , 50 , 130-115-534 , PE (APC) , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: T cell subset classification and spatial distribution in an HCC sample. (A) Bar graph quantifications of gated T cell CD3 + T cells subpopulations (CD4 + T h , CD8 + T c , FoxP3 + /CD25 + T reg ) as well as T cell differentiation subsets (T N , T SCM , T CM , T EM , T EMRA , T TE ) for HCC tumor margin and tumor core. (B) Subclassification criteria used for T cell differentiation subset gating. Detailed gating scheme see Supplementary Figures S5A–D Spatial distribution of CD3+ T cell subsets as well as CD4+ and CD8+ differentiation subsets for tumor margin (C) and tumor core (D) . Arrowheads in (C, D) highlight the spatial distribution of regulatory T cells (T reg ). (E) Violin plots demonstrating expression levels of markers used for the definition of T cell differentiation subsets in tumor margin and tumor core: CD25, FoxP3, CD45RA, CD45RO, CD27, CD95, CD11a. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: Tumor margin (ROI15) and tumor core (ROI16): 975 x 769 µm.

Article Snippet: CD25 , REA945 , 50 , 130-115-534 , PE (APC) , Miltenyi Biotec.

Techniques: Cell Differentiation, Expressing

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Spatial profiling of T cell activation and exhaustion states in an HCC sample. All images depict tumor margin (left) and tumor core (right). (A) Activation status of T cells subsets (CD69 = early, CD25 = late, HLA-DR = very late). (B) PD1 marker expression on T cells, categorized as low (0-25%), intermediate (25-75%), and high (75-100%). (C) Exhaustion levels of T cells based on the expression of PD1 (single exhaustion), PD1 plus TIM3 or LAG3 (double exhaustion), or PD1 plus TIM3 and LAG3 (triple exhaustion). Arrowheads in (A–C) highlight the ITM-proximal immune cell cluster in tumor margin and core. (D) Representation of activation/exhaustion markers for T cells (CD69, CD25, HLA-DR, PD1, LAG3, TIM3, CTLA4) as violin plots for T cell activation/exhaustion subsets. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: Tumor margin (ROI15) and tumor core (ROI16): 975 x 769 µm.

Article Snippet: CD25 , REA945 , 50 , 130-115-534 , PE (APC) , Miltenyi Biotec.

Techniques: Activation Assay, Marker, Expressing