anti-cd4 mab gk1.5  (Bio X Cell)

Journal: Veterinary Sciences

Article Title: Evaluation of PD-L1 and TIM-3 Pathways in T Cells During Experimental Bovine Leukemia Virus Infection in Sheep

doi: 10.3390/vetsci12090810

Figure Lengend Snippet: Figure 1. Cross-reactivity of anti-bovine PD-L1 and TIM-3 monoclonal antibodies in ovine PBMCs. (A) Cross-reactivity of anti-bovine PD-L1 monoclonal antibody (mAb) with fresh ovine PBMCs. The gray histogram shows fluorescence intensity of the rat IgG2a isotype control. The pink histogram shows staining with anti-bovine PD-L1 mAb. All data were gated on live IgM+ cells within PBMCs. (B–D) Cross-reactivity of anti-bovine TIM-3 mAb with fresh ovine PBMCs. The gray histograms show fluorescence intensity of the mouse IgG1 isotype control. The red histograms show staining with anti-bovine TIM-3 mAb. Histograms were gated on CD4+ (B), CD8+ (C), and γδTCR+ T cells (D). Gating strategies for the flow cytometric assays were shown in Supplementary Figure S1.

Article Snippet: Cells were then washed and stained with Alexa Fluor 647-conjugated anti-mouse IgG antibody (Thermo Fisher Scientific), Alexa Fluor 488-labeled anti-ovine CD4 mAb (17D; mouse IgG1; Washington State University Monoclonal Antibody Center, Pullman, WA, USA), phycoerythrin (PE)-conjugated anti-bovine CD8 mAb (CC63; mouse IgG1; Bio-Rad, Hercules, CA, USA), PE/cyanine7 (PE/Cy7)-conjugated anti-ovine γδTCR mAb (86D; mouse IgG1; Washington State University Monoclonal Antibody Center), and Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific) at 4 ◦C for 20 min.

Techniques: Bioprocessing, Fluorescence, Control, Staining

Journal: Veterinary Sciences

Article Title: Evaluation of PD-L1 and TIM-3 Pathways in T Cells During Experimental Bovine Leukemia Virus Infection in Sheep

doi: 10.3390/vetsci12090810

Figure Lengend Snippet: Figure 2. Blockade effects of anti-bovine PD-L1 and TIM-3 monoclonal antibodies in PBMCs of healthy sheep. (A–C) T-cell functional analysis in PBMCs from BLV-uninfected healthy sheep (n = 10) with blockade by anti-PD-L1 and anti-TIM-3 mAbs under ConA stimulation. (A) Fre- quency of activated T cells (CD25+CD69+) within CD4+ and CD8+ T-cell subsets. (B,C) Frequency of cytokine-producing activated cells (CD69+IFN-γ+ and CD69+TNF-α+) within CD4+ and CD8+ T-cell subsets. Gating strategies for the flow cytometric assays are shown in Supplementary Figures S2 and S3. Each symbol represents data from an individual animal. Red lines indicate median values. Significant differences compared between treatment groups: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells were then washed and stained with Alexa Fluor 647-conjugated anti-mouse IgG antibody (Thermo Fisher Scientific), Alexa Fluor 488-labeled anti-ovine CD4 mAb (17D; mouse IgG1; Washington State University Monoclonal Antibody Center, Pullman, WA, USA), phycoerythrin (PE)-conjugated anti-bovine CD8 mAb (CC63; mouse IgG1; Bio-Rad, Hercules, CA, USA), PE/cyanine7 (PE/Cy7)-conjugated anti-ovine γδTCR mAb (86D; mouse IgG1; Washington State University Monoclonal Antibody Center), and Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific) at 4 ◦C for 20 min.

Techniques: Bioprocessing, Functional Assay

Journal: Veterinary Sciences

Article Title: Evaluation of PD-L1 and TIM-3 Pathways in T Cells During Experimental Bovine Leukemia Virus Infection in Sheep

doi: 10.3390/vetsci12090810

Figure Lengend Snippet: Figure 4. Comparison of immunoinhibitory receptor expression between healthy and experimentally BLV-infected sheep. (A) Percentage of PD-L1+ cells among IgM+ B cells in BLV-uninfected (n = 4) and BLV-infected sheep (n = 7). (B–D) Comparison of TIM-3+ cell frequencies in CD4+, CD8+, and γδTCR+

Article Snippet: Cells were then washed and stained with Alexa Fluor 647-conjugated anti-mouse IgG antibody (Thermo Fisher Scientific), Alexa Fluor 488-labeled anti-ovine CD4 mAb (17D; mouse IgG1; Washington State University Monoclonal Antibody Center, Pullman, WA, USA), phycoerythrin (PE)-conjugated anti-bovine CD8 mAb (CC63; mouse IgG1; Bio-Rad, Hercules, CA, USA), PE/cyanine7 (PE/Cy7)-conjugated anti-ovine γδTCR mAb (86D; mouse IgG1; Washington State University Monoclonal Antibody Center), and Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific) at 4 ◦C for 20 min.

Techniques: Comparison, Expressing, Infection

Journal: Veterinary Sciences

Article Title: Evaluation of PD-L1 and TIM-3 Pathways in T Cells During Experimental Bovine Leukemia Virus Infection in Sheep

doi: 10.3390/vetsci12090810

Figure Lengend Snippet: Figure 5. T-cell activation and cytokine responses in BLV-infected sheep following immune checkpoint blockade. (A,B) Frequencies of activated CD25+CD69+ cells in CD4+ and CD8+ T-cell subsets in PBMCs from BLV-infected sheep (n = 7) following ConA stimulation with blockade by anti-PD-L1 and anti-TIM-3 mAbs. (C–F) Frequencies of cytokine-producing CD69+ cells expressing IFN-γ (C,D) or TNF-α (E,F) under the same blockade conditions. (G) IFN-γ concentration of PBMCs from BLV- infected sheep (n = 7) following ConA stimulation with blockade by anti-PD-L1 and anti-TIM-3 mAbs. Gating strategies for the flow cytometric assays are shown in Supplementary Figures S2 and S3. Each dot represents data from an individual animal. Red lines indicate median values. Significant differences compared between treatment groups: * p < 0.05, ** p < 0.01.

Article Snippet: Cells were then washed and stained with Alexa Fluor 647-conjugated anti-mouse IgG antibody (Thermo Fisher Scientific), Alexa Fluor 488-labeled anti-ovine CD4 mAb (17D; mouse IgG1; Washington State University Monoclonal Antibody Center, Pullman, WA, USA), phycoerythrin (PE)-conjugated anti-bovine CD8 mAb (CC63; mouse IgG1; Bio-Rad, Hercules, CA, USA), PE/cyanine7 (PE/Cy7)-conjugated anti-ovine γδTCR mAb (86D; mouse IgG1; Washington State University Monoclonal Antibody Center), and Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific) at 4 ◦C for 20 min.

Techniques: Activation Assay, Infection, Expressing, Concentration Assay