cd3ε pe cy5  (Elabscience Biotechnology)

Journal: Journal for Immunotherapy of Cancer

Article Title: Expression of VISTA regulated via IFN-γ governs endogenous T-cell function and exhibits correlation with the efficacy of CD19 CAR-T cell treated B-malignant mice

doi: 10.1136/jitc-2023-008364

Figure Lengend Snippet: CAR-T cell-induced tumor lysis was not sufficient to reduce VISTA expression on syngeneic activated T cell. (A) Gating strategy of flow cytometry. (B) Percentages of B-ALL cell death in vitro. B-ALL cell alone or co-cultured with purified T-cell stimulated with anti-CD3 and anti-CD28 (T activated ) in vitro were used as the control and T-cell groups, respectively. (C) The production of cytokines was evaluated in the co-culture cellular system for 6 hours. Naïve T cells were included as a control group without antibody activation (T unactivated ). (D) The representative flow cytometry histograms of VISTA levels. The T cells lacking CAR chimerism in the CAR-T groups were designated as GFP − T cells. Three independent experiments were repeated. B-ALL, B acute lymphoblastic leukemia; CAR, chimeric antigen receptor; FMO, fluorescence minus one; GFP, green fluorescent protein; VISTA, V-domain Ig suppressor of T-cell activation.

Article Snippet: Prior to flow cytometry analysis, staining was performed using the following monoclonal antibodies: CD3 FITC (fluorescein isothiocyanate) (clone17A2; BioLegend), CD3 BV650 (clone17A2; BioLegend), CD3ε PE/Cy5 (clone145-2C11; Elabscience), CD4 PE/Cyanine7 (cloneGK1.5; BioLegend), CD4 Alex Flour700 (cloneRM4-5; BioLegend); CD8a APC (allophycocyanin) (clone53-6.7; BioLegend), CD8 APC/Cyanine7 (clone53-6.7; BioLegend); CD11b PB (Pacific Blue) (cloneM1/70; BioLegend), F4/80 BV605 (cloneBM8; BioLegend), MHC-II AF700 (cloneM5/114.15.2; BioLegend), CD19 PE-cf594 (clone6D5; BioLegend), CD19 BV510 (clone6D5; BioLegend), CD11c PE-cf594 (cloneN418; BioLegend), IFN-γ PE/Cyanine7 (cloneXMG1.2; BioLegend), TNF-α APC (cloneMP6-XT22; BioLegend); Granzyme B eFlour450 (cloneM5/114.15.2; Thermo Fisher Scientific); CD62L PB (cloneMEL-1; BD Biosciences), CD44 APC/eFlour780 (cloneIM7; BD Biosciences); PD-1 PE/Cyanine7 (clone RMP1-30; BioLegend), TIGIT (Vstm3) PE (clone1G9; BioLegend); NGFR (CD271) APC (cloneME204; BioLegend), VISTA (PD-1H) PE (cloneMIH63; BioLegend), CD69 BV510 (cloneH1.2F3; BioLegend), and CD20 APC/Cyanine7 (clone SA275A11; BioLegend).

Techniques: Lysis, Expressing, Flow Cytometry, In Vitro, Cell Culture, Purification, Control, Co-Culture Assay, Activation Assay, Fluorescence

Journal: Cell Reports Medicine

Article Title: CD8 + T cell-intrinsic IL-6 signaling promotes resistance to anti-PD-L1 immunotherapy

doi: 10.1016/j.xcrm.2022.100878

Figure Lengend Snippet:

Article Snippet: Cells were then washed and stained with combinations of the following antibodies: CD45-BV510 (2 μg/mL; BD Biosciences, San Jose, CA; clone 30-F11), Thy1.2-efluor450 (2 μg/mL; eBioscience, Thermo Fisher Scientific, Waltham, MA; clone 53-2.1), Thy1.2-alexafluor700 (5 μg/mL; BioLegend, San Diego, CA; clone 53-2.1), Thy1.1-alexafluor488 (2.5 μg/mL; BioLegend, San Diego, CA; clone OX-7), CD3ε-PE/Cy5.5 (2 μg/mL; eBioscience, Thermo Fisher Scientific, Waltham, MA; clone 145-2C11), CD4-BUV395 (2 μg/mL; BD Biosciences, San Jose, CA, clone GK1.5), CD4-BV785 (2 μg/mL; BioLegend, San Diego, CA; clone GK1.5), CD8a-BB515 (2 μg/mL; BD Biosciences, San Jose, CA, clone 53-6.7), CD8a-PE (2 μg/mL; BioLegend, San Diego, CA; clone 53-6.7), CD8a-BUV737 (2 μg/mL; BD Biosciences, San Jose, CA; clone 53-6.7), CD11b-alexafluor700 (5 μg/mL; BioLegend, San Diego, CA; clone M1/70), Gr1-PE-Cy5.5 (1 μg/mL; eBioscience, Thermo Fisher Scientific, Waltham, MA; clone RB6-8C5), CD11c-PE/Dazzle594 (2 μg/mL; BioLegend, San Diego, CA; clone N418), MHCII (I-A/I-E)-FITC (2.5 μg/mL; eBioscience, Thermo Fisher Scientific, Waltham, MA; clone M5/114.15.2), CD64-PE/Cy7 (2 μg/mL; BioLegend, San Diego, CA; clone X54-5/7.1), CD169-PE/Cy7 (2 μg/mL; BioLegend, San Diego, CA; clone 3D6.112), B220-BUV737 (2 μg/mL; BD Biosciences, San Jose, CA; clone RA3-6B2), IL6R-PE (2 μg/mL; BioLegend, San Diego, CA; clone D7715A7), and gp130-PE (2 μg/mL; BioLegend, San Diego, CA).

Techniques: Recombinant, Cell Stimulation, Staining, Cell Isolation, Expressing, CRISPR, Negative Control, In Situ Hybridization, Software

Journal: Arthritis Research & Therapy

Article Title: Aryl hydrocarbon receptor signals attenuate lung fibrosis in the bleomycin-induced mouse model for pulmonary fibrosis through increase of regulatory T cells

doi: 10.1186/s13075-020-2112-7

Figure Lengend Snippet: FICZ did not affect total number of several lymphocyte subsets in the lung. BLM at 0.06 units/animal was intratracheally administered with (denoted as FICZ) or without FICZ (vehicle). Flow cytometry was used to examine single-cell suspensions extracted from lung tissues stained with anti-mouse CD45, CD3, CD4, γδ, CD8, B220, NK-1.1, and viability dye 1 week after BLM administration. Total number of CD3 + T cells ( a ), CD3 + CD4 + T cells ( b ), CD3 + CD8 + T cells ( c ), CD3 + γδ + T cells ( d ), CD3 − B220 − NK1.1 + NK cells ( e ), and B220 + B cells ( f ) was compared between the two groups. n = 5 in each group using Mann-Whitney test

Article Snippet: A single-cell suspension was stained with anti-mouse CD45 (BV510, BD Biosciences), CD3ε (PE-cy5, Thermo Fisher Scientific), CD4 (eFluor® 450, Thermo Fisher Scientific), γδTCR (FITC, Thermo Fisher Scientific), CD8a (PE-CyTM, BD Biosciences), B220 (PE, Thermo Fisher Scientific), and NK-1.1 (APC, BD Biosciences) for surface staining.

Techniques: Flow Cytometry, Staining, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Aryl hydrocarbon receptor signals attenuate lung fibrosis in the bleomycin-induced mouse model for pulmonary fibrosis through increase of regulatory T cells

doi: 10.1186/s13075-020-2112-7

Figure Lengend Snippet: FICZ increased CD4 + Foxp3 + Tregs in the lungs 1 week after BLM administration. BLM at 0.06 units/animal was intratracheally administered with (denoted as FICZ) or without FICZ (vehicle). a Representative flow cytometry plots of single-cell suspensions extracted from lung tissues stained with anti-mouse CD45, CD3, CD4, Foxp3, and viability dye 1 week after BLM administration are shown. b Summary of total number of CD4 + Foxp3 + Tregs compared between the two groups. n = 5 in each group. * p < 0.05 using Mann-Whitney test

Article Snippet: A single-cell suspension was stained with anti-mouse CD45 (BV510, BD Biosciences), CD3ε (PE-cy5, Thermo Fisher Scientific), CD4 (eFluor® 450, Thermo Fisher Scientific), γδTCR (FITC, Thermo Fisher Scientific), CD8a (PE-CyTM, BD Biosciences), B220 (PE, Thermo Fisher Scientific), and NK-1.1 (APC, BD Biosciences) for surface staining.

Techniques: Flow Cytometry, Staining, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Aryl hydrocarbon receptor signals attenuate lung fibrosis in the bleomycin-induced mouse model for pulmonary fibrosis through increase of regulatory T cells

doi: 10.1186/s13075-020-2112-7

Figure Lengend Snippet: Chronologic changes in the number of Tregs, CD4 + IFNγ + and γδ + IL-17A + T cells after BLM administration. BLM at 0.06 units/animal was intratracheally administered with (denoted as FICZ) or without FICZ (vehicle). PBS was intratracheally administered as a control of BLM. Mice were sacrificed 1 and 3 weeks after administration for flow cytometric analysis. Total number of CD3 + CD4 + T cells ( a ), CD3 + γδ + T cells ( b ), CD4 + Foxp3 + Tregs ( c ), CD4 + IFNγ + T cells ( d ), and γδ + IL-17A + T cells ( e ) were compared among the three groups. n = 5 in each group. * p < 0.05 BLM with FICZ or BLM with vehicle compared to PBS with FICZ using Mann-Whitney test. ☨ p < 0.05 BLM with FICZ compared to BLM with vehicle, and BLM with vehicle compared to PBS with FICZ using Mann-Whitney test. ‡ p < 0.05 BLM with vehicle compared to BLM with FICZ or PBS with vehicle using Mann-Whitney test. § p < 0.05 BLM with vehicle compared to BLM with FICZ, and BLM with FICZ compared to PBS with FICZ using Mann-Whitney test

Article Snippet: A single-cell suspension was stained with anti-mouse CD45 (BV510, BD Biosciences), CD3ε (PE-cy5, Thermo Fisher Scientific), CD4 (eFluor® 450, Thermo Fisher Scientific), γδTCR (FITC, Thermo Fisher Scientific), CD8a (PE-CyTM, BD Biosciences), B220 (PE, Thermo Fisher Scientific), and NK-1.1 (APC, BD Biosciences) for surface staining.

Techniques: Control, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Aryl hydrocarbon receptor signals attenuate lung fibrosis in the bleomycin-induced mouse model for pulmonary fibrosis through increase of regulatory T cells

doi: 10.1186/s13075-020-2112-7

Figure Lengend Snippet: FICZ reduced CD4 + IFNγ + and γδ + IL-17A + T cells in the lungs at 1 week. BLM at 0.06 units/animal was intratracheally administered with (denoted as FICZ) or without FICZ (vehicle). Single-cell suspensions extracted from lung tissues were stained with anti-mouse CD45, CD3, CD4, γδ, IFN-γ, IL-17A, IL-22, and viability dye 1 week after BLM administration and analyzed by flow cytometry. Summary of the number of CD4 + ( A ) and γδ + T cells ( B ) producing the cytokines IFN-γ ( a ), IL-17A ( b ), and IL-22 ( c ) was compared between the two groups. n = 5 in each group. * p < 0.05 using Mann-Whitney test

Article Snippet: A single-cell suspension was stained with anti-mouse CD45 (BV510, BD Biosciences), CD3ε (PE-cy5, Thermo Fisher Scientific), CD4 (eFluor® 450, Thermo Fisher Scientific), γδTCR (FITC, Thermo Fisher Scientific), CD8a (PE-CyTM, BD Biosciences), B220 (PE, Thermo Fisher Scientific), and NK-1.1 (APC, BD Biosciences) for surface staining.

Techniques: Staining, Flow Cytometry, MANN-WHITNEY