Journal: Frontiers in Immunology

Article Title: A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

doi: 10.3389/fimmu.2023.1276151

Figure Lengend Snippet: Upon exposure to inflammatory stimuli such as LPS, or sensitizing chemicals such as 1 chloro-2,4-dinitrobenzene (DNCB) or nickel sulfate (NiSO 4 ), keratinocytes start to secrete inflammatory cytokines such as IL-1, TNF-α and IL-18. Subsequently cutaneous dendritic cells such as Langerhans cells and dermal dendritic cells become activated and start to phagocytose haptens or exogenous particles, which is accompanied by cell maturation and the upregulation of CD54 and CD86. Finally, DCs migrate to draining lymph nodes to present the processed antigen in order to activate CD4 + T cells. Created with BioRender.com .

Article Snippet: For staining the cells were incubated in Automacs Running Buffer with the following antibodies (1:50): REA Control (S)-VioGreen (Miltenyi, #130-113-444), REA Control (S)-PE (Miltenyi, #130-113-438), REA Control (S)-APC (Miltenyi, #130-113-434); REA Control (S)-PE-Vio770, (Miltenyi, #130-113-440); CD54-APC (Miltenyi, #130-121-342); CD86-APC (Miltenyi, #130-116-161), CD14-VioGreen (Miltenyi, #130-110-525), CD11c-APC (Miltenyi, #130-113-584) for 10 minutes in the dark.

Techniques:

Journal: Frontiers in Immunology

Article Title: A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

doi: 10.3389/fimmu.2023.1276151

Figure Lengend Snippet: Surface marker expression of CD54 and CD86 (depicted as fold of induction of the percentage of all positive cells) after (A) pre-treatment of THP-1-derived iDCs and (B) topical treatment of the immune competent skin model with dexamethasone for 1 h, followed by NiSO 4 treatment for 23 h. Results were depicted as fold of induction compared to the solvent control [0.3% DMSO]. (C–E) Cytokine secretion of iDCs after 1 h dexamethasone pre-treatment, followed by 23 h of NiSO 4 exposure. Error bars indicate the standard errors of the mean (n = 3 independent experiments for (A, C) and n=4 independent experiments for (B) with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001).

Article Snippet: For staining the cells were incubated in Automacs Running Buffer with the following antibodies (1:50): REA Control (S)-VioGreen (Miltenyi, #130-113-444), REA Control (S)-PE (Miltenyi, #130-113-438), REA Control (S)-APC (Miltenyi, #130-113-434); REA Control (S)-PE-Vio770, (Miltenyi, #130-113-440); CD54-APC (Miltenyi, #130-121-342); CD86-APC (Miltenyi, #130-116-161), CD14-VioGreen (Miltenyi, #130-110-525), CD11c-APC (Miltenyi, #130-113-584) for 10 minutes in the dark.

Techniques: Marker, Expressing, Derivative Assay, Solvent, Control

Journal: International Journal of Molecular Medicine

Article Title: Anti-β 2 GPI/β 2 GPI induces neutrophil pyroptosis and thereby enhances ICAM-1 and IL-8 expression in endothelial cells

doi: 10.3892/ijmm.2022.5120

Figure Lengend Snippet: HMGB1 and IL-1β promote in vitro endothelial cell activation. (A) Representative microscopic image of endothelial cell morphology (scale bar, 50 μ m). (B) Changes in endothelial cell ICAM-1 expression over time (0-24 h) (n=3). (C) IL-8 and ICAM-1 expression levels in endothelial cells following treatment for 24 h with HMGB1 (4 ng/ml), with β-actin being used for normalization (n=4). (D) IL-8 and ICAM-1 expression levels in endothelial cells stimulated for 24 h with IL-1β (2 ng/ml), with β-actin being used for normalization (n=4). (E and F) IL-8 and ICAM-1 protein levels following (E) HMGB1 or (F) IL-1β stimulation (n=3). (G) IL-8 expression as assessed via fluorescence microscopy following a 24 h stimulation with HMGB1 (4 ng/ml) or IL-1β (2 ng/ml). (H) ICAM-1 expression was assessed via fluorescence microscopy following a 24 h stimulation with HMGB1 (4 ng/ml) or IL-1β (2 ng/ml) (scale bar, 50 μ m). Values are expressed as the mean ± standard error of the mean. * P<0.05, ** P<0.01, *** P<0.001 as indicated or vs. Ctrl. Ctrl, control; ns, no significance; HMGB1, high mobility group box protein 1; ICAM-1, intercellular adhesion molecule-1.

Article Snippet: In brief, cells were fixed for 20 min with 4% paraformaldehyde and permeabilized for 30 min with 0.2% Triton X-100 at room temperature, then blocked with 50% goat serum (Beyotime Institute of Biotechnology) in PBS for 30 min at 37°C and incubated overnight with anti-GSDMD (cat. no. 20770-1-AP; 1:100 dilution; Proteintech Group, Inc.), anti-HMGB1 (cat. no. bs-55098R; 1:50 dilution; Bioss) or anti-ICAM-1 (cat. no. WL02268) or anti-IL-8 (cat. no. WL03074; both 1:50 dilution; both from Wanleibio Co., Ltd.) at 4°C.

Techniques: In Vitro, Activation Assay, Expressing, Fluorescence, Microscopy

Journal: International Journal of Molecular Medicine

Article Title: Anti-β 2 GPI/β 2 GPI induces neutrophil pyroptosis and thereby enhances ICAM-1 and IL-8 expression in endothelial cells

doi: 10.3892/ijmm.2022.5120

Figure Lengend Snippet: Schematic overview of the proposed mechanism whereby anti-β 2 GPI/β 2 GPI interactions drive neutrophil pyroptosis and associated IL-1β and HMGB1 release, thereby causing endothelial cell activation. TLR4, Toll-like receptor 4; NLRP3, NLR family pyrin domain containing 3; anti-β2GPI, anti-β2-glycoprotein I; GSDMD, gasdermin D; PKR, double-stranded RNA-dependent protein kinase; HMGB1, high mobility group box protein 1; ICAM-1, intercellular adhesion molecule-1.

Article Snippet: In brief, cells were fixed for 20 min with 4% paraformaldehyde and permeabilized for 30 min with 0.2% Triton X-100 at room temperature, then blocked with 50% goat serum (Beyotime Institute of Biotechnology) in PBS for 30 min at 37°C and incubated overnight with anti-GSDMD (cat. no. 20770-1-AP; 1:100 dilution; Proteintech Group, Inc.), anti-HMGB1 (cat. no. bs-55098R; 1:50 dilution; Bioss) or anti-ICAM-1 (cat. no. WL02268) or anti-IL-8 (cat. no. WL03074; both 1:50 dilution; both from Wanleibio Co., Ltd.) at 4°C.

Techniques: Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Heme Oxygenase Protects against Placental Vascular Inflammation and Abortion by the Alarmin Heme in Mice

doi: 10.3390/ijms21155385

Figure Lengend Snippet: Primary antibodies used for immunohistochemical staining for IL-1β, ICAM-1, F4/80 and HO-1. Source and used concentrations of the antibodies are described.

Article Snippet: CD54 , ICAM-1 , 0.34 , Proteintech via Sanbio, Uden, The Netherlands.

Techniques: Immunohistochemical staining, Staining, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD54 , REA266 , 50 , 130-120-711 , PE , Miltenyi Biotec.

Techniques: Imaging