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Cd83 apc

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CD83-APC is a fluorochrome-conjugated antibody that binds to the CD83 cell surface antigen. CD83 is a member of the immunoglobulin superfamily and is expressed on the surface of mature dendritic cells.

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Lab products found in correlation

Hla dr percp cy5, by BioLegend (3 mentions) Cd80 pe cy7, by BioLegend (2 mentions) Human ifn gamma quantikine elisa kit, by R&D Systems (2 mentions) P irf3, by Cell Signaling Technology (2 mentions) Recombinant human il 4, by R&D Systems (2 mentions) Pd l1 bv421, by BioLegend (2 mentions) Total irf3, by Cell Signaling Technology (2 mentions) Mouse anti human cd11c pe cy7, by BioLegend (2 mentions) Epcam pe cf594, by BioLegend (2 mentions) Cd86 bv510, by BioLegend (2 mentions) Pd l2 apc, by BioLegend (2 mentions) Mouse anti human cd56 fitc, by BD (2 mentions) Rabbit anti human sting, by Cell Signaling Technology (2 mentions) Mouse anti human β actin, by Bio-Rad (2 mentions) Secondary goat anti rabbit and anti mouse antibodies, by LI COR (2 mentions) Sting primary ihc antibody, by Abcam (2 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (2 mentions) Gm csf, by R&D Systems (2 mentions) Lymphocyte isolation solution, by Dakewe (1 mentions) Lsrfortessa cell analyser, by BD (1 mentions)

9 protocols using cd83 apc

1

Analyzing Splenic Immune Cells

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The spleen was collected under aseptic conditions and prepared as a single-cell suspension using a cell strainer, and peripheral blood mononuclear cells (PBMCs) were isolated from the spleen suspensions using lymphocyte isolation solution (Dakewe Biotech, Beijing, China) according to the manufacturer′s instructions. For analysis in a flow cytometer (LSRFortessa™, BD, Franklin, NJ, USA), PBMCs were stained with APC-CD83 (BioLegend, San Diego, CA, USA). FlowJo software was used to analyze the total number of lymphocytes.
Gao Y., Cheng J., Xu X., Li X., Zhang J., Ma D., Jiang G., Liao Y., Fan S., Niu Z., Yue R., Chang P., Zeng F., Duan S., Meng Z., Xu X., Li X., Li D., Yu L., Ping L., Zhao H., Guo M., Wang L., Wang Y., Zhang Y, & Li Q. (2022). HSV-1 Infection of Epithelial Dendritic Cells Is a Critical Strategy for Interfering with Antiviral Immunity. Viruses, 14(5), 1046.
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2

Quantifying Immune Cell Activation Markers

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For detection of activated caspase-3 and viral NP protein in monocytes, the cells were collected at 12 and 24hpi and fixed with 4% paraformaldehyde, the cell membranes were permeabilized with 0.1% Triton X-100 for 5 min and then incubated with Alexa Fluor 647-conjugated anti-cleaved caspase-3 antibody (BD Biosciences, CA, USA) or FITC-conjugated anti-influenza A virus NP antibody (Abcam) for 30 min at room temperature. To determine the cell surface expression of differentiation or maturation markers, culture supernatant stimulated cells were collected at 48 or 72 h post-stimulation. The cells were stained with antibodies for 15 min at room temperature and then fixed with 4% paraformaldehyde. Brilliant Violet 421-CD80, APC-CD83, Brilliant Violet 605-CD86, APC-CD3, and corresponding isotype control antibodies (all from Biolegend, San Diego, CA, USA) were used. Stained cells were analysed on LSR Fortessa cell analyser (BD Bioscience), data was analysed using Flowjo software (TreeStar, Inc).
Lee A.C., Zhang A.J., Chu H., Li C., Zhu H., Mak W.W., Chen Y., Kok K.H., To K.K, & Yuen K.Y. (2019). H7N9 influenza A virus activation of necroptosis in human monocytes links innate and adaptive immune responses. Cell Death & Disease, 10(6), 442.
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3

Quantification of CD34+mono Cells in Peripheral Blood

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Peripheral blood samples were obtained from donors at a median of 4 days after G-CSF administration (range: 3–6). Mononuclear cells were separated by density gradient sedimentation using Lymphoprep™ (Axis-Shield PoC AS), and cryopreserved at −80 °C until use. Cells were incubated with anti-human Lin (Lineage: CD3, CD19, CD56)-FITC, CD34-PE, CD33-APC or CD83-APC, CD11b-PECy7, CD14-APCCy7 monoclonal antibodies, and 7AAD (BioLegend). Data were analysed with FACSverse and FACSuite (BD Biosciences).
CD34+mono was defined as Lin−CD34highCD14+CD11b+CD33+ cells (Fig. 1). When ≥0.1% of CD34+mono among CD34-positve cells was detected with ≥5 events, the donor was considered to be positive for CD34+mono.
Nakasone H., Kikuchi M., Kawamura K., Akahoshi Y., Sato M., Kawamura S., Yoshino N., Takeshita J., Yoshimura K., Misaki Y., Gomyo A., Tanihara A., Kusuda M., Tamaki M., Kimura S.I., Kako S, & Kanda Y. (2019). Increased CD83 expression of CD34-positive monocytes in donors during peripheral blood stem cell mobilization in humans. Scientific Reports, 9, 16499.
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4

Flow Cytometry Analysis of moDC Markers

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Treated or non-treated moDCs (5 × 104 cells/well) were incubated for 15 min at RT with specific mAbs (HLA-DR–FITC, PD-L1-PE, CD86-PerCP, CD80-PE-Cy7 and CD83-APC (Biolegend, San Diego, CA, USA) or with an isotype-matched control). The cells were acquired by flow cytometry and analyzed using FlowJo software, as previously described. The results were expressed as the fold change of the percentage of expression for each surface marker on the moDCs. The fold change was calculated for each marker (% marker expression on stimulated moDCs/% marker expression on unstimulated moDCs).
Fernandez-Prades L., Brasal-Prieto M., Alba G., Martin V., Montserrat-de la Paz S., Cejudo-Guillen M., Santa-Maria C., Dakhaoui H., Granados B., Sobrino F., Palomares F, & Lopez-Enriquez S. (2023). Sulforaphane Reduces the Chronic Inflammatory Immune Response of Human Dendritic Cells. Nutrients, 15(15), 3405.
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5

Multicolor Flow Cytometry Immunophenotyping

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The following fluorescein-labeled antibodies were used for cell surface marker analysis: CD11c-PE-cy7, CD40-APC, CD83-APC, CD80-PE-cy5, MHC-II-FITC, B7-H3-PE, B7-H4-PE, CD3-PE, CD8a-PE-Cy5, CD4-APC, CD25-PE-Cy7, and Foxp3-FITC. They were purchased for eBioscience (San Diego, CA, USA) except for CD83-APC from BioLegend (USA). For intracellular staining, Via-probe was added prior to fixation and permeabilization. Data were acquired with flow cytometer and analyzed using FlowJo software.
Yang M., Zhang F., Qin K., Wu M., Li H., Zhu H., Ning Q., Lei P, & Shen G. (2016). Glucose-Regulated Protein 78-Induced Myeloid Antigen-Presenting Cells Maintained Tolerogenic Signature upon LPS Stimulation. Frontiers in Immunology, 7, 552.
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6

Multiparameter Flow Cytometry Panel

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Mouse anti-human CD11c-PE/Cy7, EPCAM-PE/CF594, CD86-BV510, HLA-DR PerCP/Cy5.5, PD-L1-BV421, PD-L2-APC, and CD83-APC were purchased from Biolegend (San Diego, CA). Fixable Viability Dye eFluor™ 780 was purchase from eBioscience (San Diego, CA) and mouse anti-human CD56-FITC were purchased from BD Biosciences (San Jose, CA). Recombinant human IL-4 and GM-CSF were purchased from R&D systems (Minneapolis, MN) reconstituted according manufacturer instructions and kept at −80 Celsius freezer in aliquots. 2′, 3′- cGAMP sodium salt was purchased from Tocris (Bristol, UK). Cetuximab (anti-EGFR mAb, IgG1) was kindly provided by Bristol-Meyers Squibb and was used at a concentration of 10 ug/mL in all our experiments. Human IgG1 Kappa-LE/AF was purchased from Sourthern Biotech (Birmingham, AL). Western blot antibodies included rabbit anti-human STING, pIRF3, and total IRF3 were purchased from Cell Signaling (Danvers, MA). Mouse anti-human β-actin was purchased from Bio Rad (Hercules, CA). Secondary goat anti-rabbit and anti-mouse antibodies were purchased from LI-COR biosciences (Lincoln, NE). STING primary IHC antibody was purchased from Abcam (Cambridge, MA). Human IFN-gamma Quantikine ELISA Kit was purchased from R&D systems (Minneapolis, MN).
Lu S., Concha-Benavente F., Shayan G., Srivastava R.M., Gibson S.P., Wang L., Gooding W.E, & Ferris R.L. (2018). STING activation enhances cetuximab-mediated NK cell activation and DC maturation and correlates with HPV+ status in head and neck cancer. Oral oncology, 78, 186-193.
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7

Immunophenotyping of Dendritic Cells

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Antibodies and other reagents used were fixable viability dye e-flour 450, Annexin V FITC (ebioscience); anti-human CD86-FITC, CD80-FITC, Propidium Iodide (PI) (BD Pharmingen, San Jose, CA); anti-human CD11c PE, HLA-DR PerCP, CD80-PE (Becton Dickinson, San Jose, CA); CD282-FITC, CD284-PE, Donkey anti-rabbit-FITC, CD86-PE, CD83-APC, CD11c-PE-Cy7, CD83-FITC, CD14-APC, and 7 aminoactinomycin D (7AAD (Biolegend, San Diego, CA). Data acquisition was on a Miltenyi MacsQuant flow cytometer (Miltenyi Biotech, San Diego, CA) on a log scale and analysis was performed with FlowJo 7.6.5 software (Tree Star, Ashland, OR).
Nicholas D.A., Zhang K., Hung C., Glasgow S., Aruni A.W., Unternaehrer J., Payne K.J., Langridge W.H, & De Leon M. (2017). Palmitic acid is a toll-like receptor 4 ligand that induces human dendritic cell secretion of IL-1β. PLoS ONE, 12(5), e0176793.
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8

Multiparameter Flow Cytometry Panel

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Mouse anti-human CD11c-PE/Cy7, EPCAM-PE/CF594, CD86-BV510, HLA-DR PerCP/Cy5.5, PD-L1-BV421, PD-L2-APC, and CD83-APC were purchased from Biolegend (San Diego, CA). Fixable Viability Dye eFluor™ 780 was purchase from eBioscience (San Diego, CA) and mouse anti-human CD56-FITC were purchased from BD Biosciences (San Jose, CA). Recombinant human IL-4 and GM-CSF were purchased from R&D systems (Minneapolis, MN) reconstituted according manufacturer instructions and kept at −80 Celsius freezer in aliquots. 2′, 3′- cGAMP sodium salt was purchased from Tocris (Bristol, UK). Cetuximab (anti-EGFR mAb, IgG1) was kindly provided by Bristol-Meyers Squibb and was used at a concentration of 10 ug/mL in all our experiments. Human IgG1 Kappa-LE/AF was purchased from Sourthern Biotech (Birmingham, AL). Western blot antibodies included rabbit anti-human STING, pIRF3, and total IRF3 were purchased from Cell Signaling (Danvers, MA). Mouse anti-human β-actin was purchased from Bio Rad (Hercules, CA). Secondary goat anti-rabbit and anti-mouse antibodies were purchased from LI-COR biosciences (Lincoln, NE). STING primary IHC antibody was purchased from Abcam (Cambridge, MA). Human IFN-gamma Quantikine ELISA Kit was purchased from R&D systems (Minneapolis, MN).
Lu S., Concha-Benavente F., Shayan G., Srivastava R.M., Gibson S.P., Wang L., Gooding W.E, & Ferris R.L. (2018). STING activation enhances cetuximab-mediated NK cell activation and DC maturation and correlates with HPV+ status in head and neck cancer. Oral oncology, 78, 186-193.
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9

Multiparametric flow cytometry of dendritic cells

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Immature and mature DCs were stained with HLA-DR PerCP/Cy5.5, CD40 FITC, CCR7 PE, CD80 PE/CY7, CD86 PE-Dazzle 594 and CD83 APC (Biolegend, USA). The cells were acquired using a LSRII flow cytometer (Beckton Dickinson, USA) and analysed using FloJo software (version 10.1; Treestar, USA). Dead cells were gated out of the scatter plots prior to analysis and negative gates were set using mean fluorescence one (MFO) controls.
Tomasicchio M., Semple L., Esmail A., Meldau R., Randall P., Pooran A., Davids M., Cairncross L., Anderson D., Downs J., Malherbe F., Novitzky N., Panieri E., Oelofse S., Londt R., Naiker T, & Dheda K. (2018). An autologous dendritic cell vaccine polarizes a Th-1 response which is tumoricidal to patient-derived breast cancer cells. Cancer Immunology, Immunotherapy, 68(1), 71-83.
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