anti arginase1 apc  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: Establishment and Characterization of Free-Floating 3D Macrophage Programming Model in the Presence of Cancer Cell Spheroids

doi: 10.3390/ijms241310763

Figure Lengend Snippet: Summary of changes in cell surface markers and secretome profile of macrophages cultivated in the presence of cancer cell spheroids compared to the same macrophages. Markers whose changes have been detected for four or more of the studied phenotypes (groups) are highlighted in bold. GM-CSF-induced chemokines are highlighted in blue. The decrease in 4T1 cell receptor CCR4, CXCR4, and CXCR7 ligands is highlighted in orange. ( w / o )—cells incubated washed out activating factors (without IFNγ/Pam3SCK4, or IL-4), ND—changes not detected, ↑ —upregulated, ↓ —downregulated.

Article Snippet: The suspension was mixed and incubated for 15 min at RT, and then 150 μL of permeabilization reagent and anti-Arginase1-APC (Cat. No 17-3697-82; Invitrogen) antibodies were added in appropriate amounts.

Techniques: Incubation

Journal: iScience

Article Title: βig-h3-structured collagen alters macrophage phenotype and function in pancreatic cancer

doi: 10.1016/j.isci.2022.103758

Figure Lengend Snippet: rβig-h3-structured collagen I fibers modulate the phenotype and function of macrophages (A) Representative photographs of immunofluorescence staining of pancreata from three-months-old KC mice stained for βig-h3 (green) and CD206 (red). Nuclear counterstaining in DAPI (blue). (B) Representative photographs of βig-h3, F4/80, CD206, and Arg1 staining on serial section of pancreata from three-months-old KC mice. (C) FACS analysis of BMMCs cultured alone or on collagen I structured in the absence (BMMCs + Col I) or presence of rβig-h3 (BMMCs + Col I rβig-h3 ). Representative data of three independent experiments with three in vitro replicates per group are shown. ∗p<0.05, ∗∗p<0.01. Error bars 50 μm, 25 μm.

Article Snippet: The following monoclonal Abs were used in flow cytometry: anti-F4/80 Pe-Cy7 (clone BM8-Biolegend, 123114) or PerCP Cy5.5 (clone BM8-Biolegend, 123128), anti-MHCII PE (clone M5/114.15.2- eBioscience, 12-5321-81), anti-CD206 BV650 (clone C068C2-Biolegend, 141723), anti-CD86 BV605 (clone GL-1- Biolegend, 105029), anti-CD80 APC-Cy7 (clone 16-10A1-Biolegend, 104729), anti-CSFRI AF488 (clone AFS98- Biolegend, 135511), anti-INOS PE (clone CXNFT, Life Techno, 12-5920-82), anti-Arginase1 APC (RD, IC5868A), anti-TGF-β B421 (clone TW7-16B4-Biolegend, 141408) and anti-TNFα BV605 (clone MP6-XT22-Biolegend, 506329).

Techniques: Immunofluorescence, Staining, Cell Culture, In Vitro

Journal: iScience

Article Title: βig-h3-structured collagen alters macrophage phenotype and function in pancreatic cancer

doi: 10.1016/j.isci.2022.103758

Figure Lengend Snippet: βig-h3 Ab depletion in vivo reprograms macrophage phenotype in the tumor microenvironment (A) Experimental setting. (B) Tumor weight of mice treated in A. (C-F) FACS analysis of the percentages of CD45 + (C), F4/80 + CD45 + (D), CD206 + F4/80 + (E), and Arg1 + F4/80 + (F) cells collected in A. ∗p<0.05, ∗∗p<0.01. Representative of two independent experiments with fivemice per group.

Article Snippet: The following monoclonal Abs were used in flow cytometry: anti-F4/80 Pe-Cy7 (clone BM8-Biolegend, 123114) or PerCP Cy5.5 (clone BM8-Biolegend, 123128), anti-MHCII PE (clone M5/114.15.2- eBioscience, 12-5321-81), anti-CD206 BV650 (clone C068C2-Biolegend, 141723), anti-CD86 BV605 (clone GL-1- Biolegend, 105029), anti-CD80 APC-Cy7 (clone 16-10A1-Biolegend, 104729), anti-CSFRI AF488 (clone AFS98- Biolegend, 135511), anti-INOS PE (clone CXNFT, Life Techno, 12-5920-82), anti-Arginase1 APC (RD, IC5868A), anti-TGF-β B421 (clone TW7-16B4-Biolegend, 141408) and anti-TNFα BV605 (clone MP6-XT22-Biolegend, 506329).

Techniques: In Vivo

Journal: iScience

Article Title: βig-h3-structured collagen alters macrophage phenotype and function in pancreatic cancer

doi: 10.1016/j.isci.2022.103758

Figure Lengend Snippet:

Article Snippet: The following monoclonal Abs were used in flow cytometry: anti-F4/80 Pe-Cy7 (clone BM8-Biolegend, 123114) or PerCP Cy5.5 (clone BM8-Biolegend, 123128), anti-MHCII PE (clone M5/114.15.2- eBioscience, 12-5321-81), anti-CD206 BV650 (clone C068C2-Biolegend, 141723), anti-CD86 BV605 (clone GL-1- Biolegend, 105029), anti-CD80 APC-Cy7 (clone 16-10A1-Biolegend, 104729), anti-CSFRI AF488 (clone AFS98- Biolegend, 135511), anti-INOS PE (clone CXNFT, Life Techno, 12-5920-82), anti-Arginase1 APC (RD, IC5868A), anti-TGF-β B421 (clone TW7-16B4-Biolegend, 141408) and anti-TNFα BV605 (clone MP6-XT22-Biolegend, 506329).

Techniques: Recombinant, Modification, Plasmid Preparation, Activation Assay, Concentration Assay, Software

Journal: Oncoimmunology

Article Title: Compensatory upregulation of PD-1, LAG-3, and CTLA-4 limits the efficacy of single-agent checkpoint blockade in metastatic ovarian cancer

doi: 10.1080/2162402X.2016.1249561

Figure Lengend Snippet: Effects of combinatorial blockade of PD-1, CTLA-4, and LAG-3 on Treg and MDSC population in ovarian TME. (A) Dual and triple antibody blockade treatment decreased the frequency of CD4 + CD25 + FoxP3 + cells at early time point of ovarian tumor progression. TALs and TILs were isolated as described above and stained for the expression of CD4 + , CD25, and FoxP3 protein and analyzed using flow cytometry. (B) Arginase 1 (ARG1)-expressing population was increased in the ovarian TME as compared with that in spleen. Splenocytes and TALs were isolated as described in Fig. 5 . Frequency of ARG1 + population in the CD45 + CD11b + cells was determined based on the gating shown in Fig. S3. (C–E) Various checkpoint blockade combinations reduced the frequencies of ARG1 + -MDSCs. The frequency of ARG1 + -Ly6G + 6C + cells was significantly reduced by PD-1/CTLA-4 dual blockade (C). All three dual blockades of PD-1/LAG-3, PD-1/CTLA-4, and LAG-3/CTLA-4 significantly reduced the frequency of ARG1 + -Ly6G − 6C + (D) and ARG1 + -F4/80 + cells (E). Data were obtained from 5 mice per group, analyzed using GraphPad Prism 6, and are representative of two independent experiments. Error bars represent SD. Statistical significance was determined by Student's t -test. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The following antibodies were used for human samples: Alexafluo-700-CD8a (Biolegend, 300920), FITC-CD4 + (eBioscience, 25-0041-82 ), Brilliant violet-421-PD-1 (Biolegend, 329919), phycoerythrin (PE)-LAG-3 (R&D systems, FAB2319P), APC-TIM-3 (Biolegend, 345011), and PE-Cy7-CTLA-4 (Biolegend, 349913), The following antibodies were used for mouse study: PE-LAG-3 (Biolegend, 125208), Brilliant violet-421-PD-1 (Biolegend, 135218), Alexafluor-700-CD8a (Biolegend, 100730), PE-Cy7-CD4 (eBioscience, 25-0041-82), APC-TIM-3 (eBioscience, 17-5871-82), and FITC-CTLA-4 (Southern Biotechnology, 1790-02), PE-CD160 (Biolegend, 143004), FITC-CD244 (2B4, eBioscience, 11-2441-82), APC-BTLA (eBioscience, 17-5956-80), FITC-KLRG1 (eBioscience, 11-5893-82), PE-Cy7-TNF-alpha (BD Biosciences, 557644), APC-IFNγ (BD Biosciences, 557644), FITC-IL2 (BD Biosciences, 554427), APC-CD25 (eBioscience, 17-0251-82) PE-FOXP3 (eBioscience, 12-5773-82), Live-dead stain (Near IR fluorescent reactive dye; Molecular probe, L10119), V450-CD45 (BD Biosciences, 560501), CD11b (BD Biosciences, 557397), FITC-Ly-6G (BD Biosciences, 551460), Alexa Fluor 700-Ly-6C (BD Biosciences 581237), PerCP-Cyanine5.5-F4/80 Antigen (Biolegend, 123128), anti-human/mouse Arginase1 APC-conjugated (R&D System, IC5868A).

Techniques: Isolation, Staining, Expressing, Flow Cytometry