anti ovine cd4 mab  (Bio-Rad)

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

Journal: Cell Reports Medicine

Article Title: Fc-optimized anti-CTLA-4 antibodies increase tumor-associated high endothelial venules and sensitize refractory tumors to PD-1 blockade

doi: 10.1016/j.xcrm.2025.102141

Figure Lengend Snippet: Anti-CTLA-4 antibodies remodel tumor vasculature and increase TA-HEVs through Fc-dependent mechanisms (A) Treatment schedule. i.p., intraperitoneal; IF, immunofluorescence; control, isotype control antibodies. (B) Mean tumor growth ( n = 9 mice per group; data pooled from two independent experiments). (C) Gating strategy for CD45 − CD31 high endothelial cells and MECA-79 + TA-HECs. (D and E) Frequencies of CD45 − CD31 high endothelial cells and TA-HECs. Each symbol represents an individual mouse ( n = 9, two independent experiments). (F and G) Mean fluorescence intensity (MFI) of MECA-79 and CD62P in TA-HECs, quantified by flow cytometry. Each symbol represents an individual mouse ( n = 9, two independent experiments). (H and I) Immunofluorescence of MCA prog tumor (day 12) following treatment with 9D9 mouse anti-CTLA-4 antibody. Selected markers are presented. Scale bars: 100 μm. (J) Schematics of the two distinct 9D9 anti-CTLA-4 antibodies used in the study. (K and L) Mean tumor growth and tumor weights in individual mice ( n = 15 mice per group; data pooled from three independent experiments). (M–O) Frequencies of CD45 − CD31 high endothelial cells, TA-HECs, and MECA-79 + CD62P + TA-HECs. Each symbol represents an individual mouse ( n = 15, three independent experiments). Representative dot plots of TA-HECs are shown (N). (P and Q) MFI of MECA-79 and CD62P in TA-HECs, quantified by flow cytometry. Each symbol represents an individual mouse ( n = 15, three independent experiments). (R) Numbers of tumor-infiltrating CD4 + and CD8 + T cells. Each symbol represents an individual mouse ( n = 15, three independent experiments). Data are shown as mean ± SEM. All p values were determined by one-way ANOVA with Tukey’s multiple comparison test, except for (B) and (K) for which p values were determined by two-way ANOVA with Dunnett’s multiple comparison test. See also and .

Article Snippet: InVivo MAb Rat anti-Mouse CD4 (clone GK1.5) , Bio X Cell , Cat# BE0003-1; RRID: AB_1107636.

Techniques: Immunofluorescence, Control, Fluorescence, Flow Cytometry, Comparison

Journal: Cell Reports Medicine

Article Title: Fc-optimized anti-CTLA-4 antibodies increase tumor-associated high endothelial venules and sensitize refractory tumors to PD-1 blockade

doi: 10.1016/j.xcrm.2025.102141

Figure Lengend Snippet: Tbet + ICOS + Th1-like CD4 + T cells correlate with TA-HEVs during anti-CTLA-4 therapy (A–C) Frequency of FOXP3 + Tregs and numbers of FOXP3 − Tconv and FOXP3 + Tregs in tumors following indicated treatments. Representative dot plots of FOXP3 + Tregs are shown (A). Each symbol represents an individual mouse ( n = 15, three independent experiments). (D) Ratio of Tconv to Treg cell numbers in tumors following indicated treatments. Each symbol represents an individual mouse ( n = 15, three independent experiments). (E) Mean fluorescence intensity (MFI) of selected markers in Tregs, quantified by flow cytometry. Each symbol represents an individual mouse (FOXP3, Tbet, PD-1, and ICOS, n = 15, three independent experiments; CTLA-4 and CD39, n = 7–8, two independent experiments). (F) Frequency of CTLA-4 + CD25 + Tregs in tumors following indicated treatments. Representative dot plots are shown. Each symbol represents an individual mouse ( n = 7–8, two independent experiments). (G) MFI of selected markers in Tconv, quantified by flow cytometry. Each symbol represents an individual mouse ( n = 15, three independent experiments). (H and I) Frequency and number of Tbet + ICOS + Th1-like CD4 + T cells in tumors following indicated treatments. Representative dot plots are shown (H). Each symbol represents an individual mouse ( n = 15, three independent experiments). (J) Frequency of Ki67 high cells in Tbet + ICOS + Th1-like CD4 + T cells in tumors following indicated treatments. Each symbol represents an individual mouse ( n = 15, three independent experiments). (K and L) Correlation analysis between the number of tumor-infiltrating Tbet + ICOS + Th1-like CD4 + T cells and the frequencies of CD45 − CD31 high endothelial cells or MECA-79 + TA-HECs. Each symbol represents an individual mouse, and simple linear regression lines are shown in red (data pooled from six independent experiments; n = 30 mice treated with 9D9 or 9H10 anti-CTLA-4 antibodies). p values and Pearson’s r were determined by Pearson correlation test. (M) Histograms showing expression of PD-1 in Tbet − ICOS − (DN), Tbet − ICOS + (SP), and Tbet + ICOS + (DP) FOXP3 − Tconv from MCA prog tumors (day 12) following treatment with 9D9 anti-CTLA-4 antibody, quantified by flow cytometry. MFI of PD-1 is quantified in indicated subsets. Each symbol represents an individual mouse ( n = 15, three independent experiments). (N) Frequency of IFNγ + cells in CD44 + PD-1 + CD4 + T cells stimulated ex vivo , following indicated treatments. Each symbol represents an individual mouse ( n = 9, two independent experiments). Data are shown as mean ± SEM. All p values were determined by one-way ANOVA with Tukey’s multiple comparison test except for graphs in (E)–(G) and (N) for which p values were determined by unpaired two-tailed Student’s t test. See also .

Article Snippet: InVivo MAb Rat anti-Mouse CD4 (clone GK1.5) , Bio X Cell , Cat# BE0003-1; RRID: AB_1107636.

Techniques: Fluorescence, Flow Cytometry, Expressing, Ex Vivo, Comparison, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Fc-optimized anti-CTLA-4 antibodies increase tumor-associated high endothelial venules and sensitize refractory tumors to PD-1 blockade

doi: 10.1016/j.xcrm.2025.102141

Figure Lengend Snippet: CD4 + T cells are important for the increase of TA-HEVs during anti-CTLA-4 therapy (A) Treatment schedule. i.p., intraperitoneal; control, isotype control antibodies. (B and C) Frequencies of CD45 − CD31 high endothelial cells and MECA-79 + TA-HECs. Each symbol represents an individual mouse ( n = 8–9, two independent experiments). (D and E) Number of tumor-infiltrating CD8 + T cells and frequency of Ki67 high cells in tumor-infiltrating CD8 + T cells. Each symbol represents an individual mouse ( n = 8–9, two independent experiments). (F and G) Frequency of PD-1 high cells in tumor-infiltrating CD44 + CD8 + T cells following indicated treatments, and mean fluorescence intensity (MFI) of PD-1 in the same subset, quantified by flow cytometry. Representative dot plots are shown (F). Each symbol represents an individual mouse ( n = 8–9, two independent experiments). (H and I) Frequency and number of GzmB + cells in tumor-infiltrating CD44 + PD-1 + CD8 + T cells. Representative dot plots are shown (H). Each symbol represents an individual mouse ( n = 8–9, two independent experiments). (J) Treatment schedule. i.p., intraperitoneal; control, isotype control antibodies. (K–Q) Frequencies of CD45 − CD31 high endothelial cells, TA-HECs, MECA-79 + CD62P + TA-HECs, number of TA-HECs, and MFI of MECA-79 and CD62P in TA-HECs, following indicated treatments, quantified by flow cytometry. Each symbol represents an individual mouse ( n = 10, two independent experiments). (R and S) Numbers of tumor-infiltrating CD4 + and CD8 + T cells, and frequency of Ki67 high cells in tumor-infiltrating CD8 + T cells. Each symbol represents an individual mouse ( n = 10, two independent experiments). Data are shown as mean ± SEM. All p values were determined by one-way ANOVA with Tukey’s multiple comparison test except for graphs in (K)–(S) for which p values were determined by unpaired two-tailed Student’s t test. See also and .

Article Snippet: InVivo MAb Rat anti-Mouse CD4 (clone GK1.5) , Bio X Cell , Cat# BE0003-1; RRID: AB_1107636.

Techniques: Control, Fluorescence, Flow Cytometry, Comparison, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Fc-optimized anti-CTLA-4 antibodies increase tumor-associated high endothelial venules and sensitize refractory tumors to PD-1 blockade

doi: 10.1016/j.xcrm.2025.102141

Figure Lengend Snippet: Fc-optimized ipilimumab combined with anti-PD-1 increases TA-HEVs and melanoma tumor control in humanized mice (A) Treatment schedule. i.p., intraperitoneal; control, isotype control antibodies. (B–D) Mean tumor growth, Kaplan-Meier survival curves, and individual tumor growth. Data were obtained from two independent experiments (control, n = 12 mice; anti-PD-1, n = 10 mice; ipilimumab-GAALIE, n = 12 mice; ipilimumab-GAALIE + anti-PD-1, n = 12 mice). (E) Treatment schedule. i.p., intraperitoneal; IF, immunofluorescence; control, isotype control antibodies. (F–H) Immunofluorescence of B16F10 tumor (day 15) following treatment with ipilimumab-GAALIE + anti-PD-1 antibodies. Selected markers are presented. Inset in (G) and (H) show higher magnifications of boxed areas. Scale bars: 500 μm (F), 50 μm (G left, H left, and H right), 20 μm (G right). (I and J) Frequencies of TA-HECs and MECA-79 + CD62P + TA-HECs. Each symbol represents an individual mouse ( n = 9–10, two independent experiments). (K and L) Frequency and numbers of Tbet + ICOS + Th1-like CD4 + T cells in tumors. Representative dot plots are shown (K). Each symbol represents an individual mouse ( n = 9–10, two independent experiments). (M) Number of tumor-infiltrating CD8 + T cells. Each symbol represents an individual mouse ( n = 9–10, two independent experiments). Data are shown as mean ± SEM. All p values were determined by unpaired two-tailed Student’s t test except for (B) for which p values were determined by two-way ANOVA with Dunnett’s multiple comparison test, and for graph in (C) for which p values were determined by log rank test.

Article Snippet: InVivo MAb Rat anti-Mouse CD4 (clone GK1.5) , Bio X Cell , Cat# BE0003-1; RRID: AB_1107636.

Techniques: Control, Immunofluorescence, Two Tailed Test, Comparison

Journal: Cell reports

Article Title: Acetyl-CoA synthesis in the skin is a key determinant of systemic lipid homeostasis

doi: 10.1016/j.celrep.2025.115284

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: InVivo MAb rat monoclonal anti-mouse CD4 , Bio X Cell , Cat#BE0003-1; RRID: AB_1107642.

Techniques: Western Blot, Staining, Control, Recombinant, Protease Inhibitor, Mass Spectrometry, Bicinchoninic Acid Protein Assay, Acid Assay, Colorimetric Assay, Software, Quantitative Proteomics

Journal: Dose-Response

Article Title: Comparative Analysis of Concurrent vs Sequential Administration of anti-PD-1 Following Thoracic Radiotherapy: Impact on Lung Tissue Damage

doi: 10.1177/15593258251322324

Figure Lengend Snippet: Recruitment of inflammatory infiltration cells in the lung tissue. (A) T lymphocytes infiltrated lung tissue in different groups. Green fluorescence: CD4 + T cells, red fluorescence: CD8 + T cells, and DAPI: blue fluorescence. (B) Infiltrating macrophages in lung tissue in different groups. (C) Infiltrating neutrophils in lung tissue in different group. * P < .05, ** P < .01.

Article Snippet: The collected BALF cells were stained 30 min at 4°C in 0.2% bovine serum albumin in PBS containing 1:100 dilution of the following antibodies: APC-Cy7 anti-mouse CD45 (BD Biosciences, cat. no. 557659), APC anti-mouse CD3 (CST, cat. no. 24265), FITC anti-mouse CD4 (CST, cat. no. 96127), BV605 anti-mouse CD8 (BD Biosciences, cat. no. 563152), PE anti-mouse CD11b (CST, cat. no. 24965), BV421 anti-mouse F4/80 (BD Biosciences, cat. no. 565411), PerCP-cy5.5 anti-mouse ly6G (CST, cat. no. 63460).

Techniques: Fluorescence