Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were stripped and re-probed for total AKT, NFκB p65, p38 MAPK, STAT1, STAT3, and β-actin (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA) as a loading control.

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were blocked with 5 % non-fat milk in TBS-T and incubated overnight at 4 °C with primary antibodies against phospho-AKT (Thr308), phospho-NFκB p65 (Ser536), phospho-p38 MAPK (Thr180/Tyr182), phospho-STAT1 (Tyr701), and phospho-STAT3 (Tyr705) (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were stripped and re-probed for total AKT, NFκB p65, p38 MAPK, STAT1, STAT3, and β-actin (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA) as a loading control.

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Cells

Article Title: Inflammatory Signaling and Endothelial Activation Drive Thrombosis in Hodgkin and Non-Hodgkin Lymphoma

doi: 10.3390/cells15080667

Figure Lengend Snippet: Activation of inflammatory signaling pathways in lymphomas. Peripheral blood mononuclear cells (MNCs) were isolated from patients with diffuse large B-cell lymphoma (DLBCL) with ( n = 15–19) or without thrombosis ( n = 9–11), follicular lymphoma (FL, n = 9–11) and Hodgkin lymphoma (HL, n = 10–12) and activation of ( A ) NFκB, ( B ) STAT3 and ( C ) p38 MAPK signaling pathways were analyzed relative to those in healthy volunteers (control, n = 10). ( D ) Kaplan–Meier curve presenting thrombosis-free survival in DLBCL patients stratified by pSTAT3/STAT3 ratio in MNC. Tick marks indicate censored observations. Representative immunoblot images corresponding to the graphs, showing phosphorylated signaling proteins and their respective total protein levels (used for ratio quantification). Values are mean ± SEM. * p < 0.05, ** p < 0.01 vs. control or as indicated.

Article Snippet: Signal has been detected using antibodies directed against phospho-NFκB (Ser536) (3031, Cell Signaling, Danvers, MA, USA), total NFκB (E-AB-22066, Elabscience, Houston, TX, USA), phospho-p38 (Tyr180/Tyr182) (E-AB-21027, Elabscience), total p38 (E-AB-66279, Elabscience), STAT3 (Cell Signaling, 9132, Danvers, MA, USA) and pSTAT3 Tyr705 (Cell Signaling, 9131S) and incubated overnight at 4 °C.

Techniques: Activation Assay, Protein-Protein interactions, Isolation, Control, Western Blot

Journal: Cells

Article Title: Inflammatory Signaling and Endothelial Activation Drive Thrombosis in Hodgkin and Non-Hodgkin Lymphoma

doi: 10.3390/cells15080667

Figure Lengend Snippet: Inflammatory stimulation of platelets aggregation in lymphoma. ( A ) Schematic representation of the experimental setup for the co-culture of HMEC-1 endothelial cells, lymphoma-derived mononuclear cells (MNCs), and platelets during 24 h of incubation using an automated microfluidic flow chamber system. HMEC-1 cells were pretreated with tumor necrosis factor alpha (TNFα, 5 ng/mL) and interleukin-6 (IL-6, 20 ng/mL) for 24 h before the introduction of labeled MNCs and platelets. Open boxes represent control samples, gray boxes indicate TNF-α–treated samples, and black boxes correspond to IL-6–treated samples. After 30 min of flow, immunofluorescence staining was performed, and the number and area of CD41a+ platelet aggregates were quantified. ( B ) Quantity and ( C ) area of CD41a+ platelet aggregates from DLBCL ( n = 4), FL ( n = 3) and HL ( n = 5) patients co-cultured with HMEC-1 cells pretreated with TNFα, IL-6 or vehicle. ( D ) Immunofluorescence images show platelet aggregates stained with CD41a antibody (white arrows), MNC labeled with CFSE (green), and HMEC-1 nuclei stained with DAPI (blue) in the microfluidic flow chamber. ( E ) Schematic diagram represents our two-hit model of inflammation-driven thrombosis development in lymphoma: NFκB and p38 activation drives endothelial activation across all lymphoma subtypes, leading to an increased thrombotic risk. The critical molecular event for thrombosis occurrence in DLBCL is IL6/STAT3-driven platelet aggregation combined with loss of NO-mediated vascular protection. Values are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. corresponding non-treated lymphomas (*) and # p < 0.05, ## p < 0.01, ### p < 0.001 vs. DLBCL (#). Abbreviations: TEM, transendothelial migration.

Article Snippet: Signal has been detected using antibodies directed against phospho-NFκB (Ser536) (3031, Cell Signaling, Danvers, MA, USA), total NFκB (E-AB-22066, Elabscience, Houston, TX, USA), phospho-p38 (Tyr180/Tyr182) (E-AB-21027, Elabscience), total p38 (E-AB-66279, Elabscience), STAT3 (Cell Signaling, 9132, Danvers, MA, USA) and pSTAT3 Tyr705 (Cell Signaling, 9131S) and incubated overnight at 4 °C.

Techniques: Co-Culture Assay, Derivative Assay, Incubation, Labeling, Control, Immunofluorescence, Staining, Cell Culture, Activation Assay, Migration