Journal: Frontiers in Immunology

Article Title: Lung epithelial and alveolar macrophage-like cell interactions significantly modify innate responses to bacterial endotoxin with the involvement of direct cellular contacts, TNF-α, ICAM1 and MCP-1

doi: 10.3389/fimmu.2025.1715943

Figure Lengend Snippet: MLE-12 cells express increased levels of ICAM-1 when in co-culture with MPI cells or in medium conditioned with MPI cell secretions, which also induce apoptosis. (A) ICAM-1 expression levels in hCD 68 GFP MPI and MLE-12 cell cultures and co-cultures when stimulated with LPS. Cells were stimulated with LPS for 24h and stained for ICAM-1 expression, with cell populations being differentiated by constitutive GFP expression of MPI cells. (B) ICAM-1 expression levels in MLE-12 exposed to media from MPI or LPS-stimulated MPI cells (Activated MPI media) or to regular MLE culture DMEM media. Polymyxin B was added to all conditions, including DMEM and MPI media, to neutralize residual LPS and ensure proper normalization across treatments. All conditions contained polymyxin B to ensure correct normalization across treatments. (C) induction of apoptosis in MLE-12 cells when exposed to media from MPI or LPS stimulated MPI cells. MLE-12 cells were incubated in media conditioned by MPI cells, naive or stimulated with 100 ng/mL LPS, for 24h and stained for ICAM-1 expression. Cell populations were differentiated by GFP expression of hCD 68 GFP MPI cells. Histograms are representative of three independent experiments and bar charts are expressed as mean ± SD for three biological replicates. Pairwise comparisons were determined by Prism software using the One-Way ANOVA followed by Dunnett’s post hoc test. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001. ns was used to indicate when the differences were found to be non-significant.

Article Snippet: For ICAM-1 neutralization, MLE-12 monolayers were incubated with 5 μg/mL of ICAM-1 blocking antibody (Mouse ICAM-1/CD54, R&D Systems) for 1h prior to addition of MPI cells, as previously reported ( ).

Techniques: Co-Culture Assay, Expressing, Staining, Incubation, Software

Journal: Frontiers in Immunology

Article Title: Lung epithelial and alveolar macrophage-like cell interactions significantly modify innate responses to bacterial endotoxin with the involvement of direct cellular contacts, TNF-α, ICAM1 and MCP-1

doi: 10.3389/fimmu.2025.1715943

Figure Lengend Snippet: TNF-α secreted from MPI cells induces ICAM-1 expression and apoptosis in MLE-12 cells. Effect of recombinant TNF-α on ICAM-1 expression (A) and apoptosis (B) in MLE-12 cells. MLE-12 cells were incubated with increasing concentrations of recombinant TNF-α for 24h. Effect of TNF-α inhibition in MPI secretions on ICAM-1 expression (C) and apoptosis (D) in MLE-12 cells. Polymyxin B was added to all conditions, including MPI media, to neutralize residual LPS and ensure proper normalization across treatments. Histograms are representative of three independent experiments and bar charts are expressed as mean ± SD for three experimental replicates. Multiple pairwise comparisons were performed by Prism software using One-Way ANOVA followed by Dunnett’s post hoc test. **p-value < 0.01; ***p-value < 0.001. ns was indicated when the differences were found to be non-significant.

Article Snippet: For ICAM-1 neutralization, MLE-12 monolayers were incubated with 5 μg/mL of ICAM-1 blocking antibody (Mouse ICAM-1/CD54, R&D Systems) for 1h prior to addition of MPI cells, as previously reported ( ).

Techniques: Expressing, Recombinant, Incubation, Inhibition, Software

Journal: Frontiers in Immunology

Article Title: Lung epithelial and alveolar macrophage-like cell interactions significantly modify innate responses to bacterial endotoxin with the involvement of direct cellular contacts, TNF-α, ICAM1 and MCP-1

doi: 10.3389/fimmu.2025.1715943

Figure Lengend Snippet: MLE-12 cells exposed to LPS-activated MPI cells supernatant can in turn induce a chemokine response in naïve MPI cells, even in the absence of other inflammatory stimulants, with MCP-1 being dependent on ICAM-1. Induction of MCP-1, MIP-2 and IP-10 in co-cultures of naïve MPI cells with MLE-12 cells exposed to LPS-activated MPI cells supernatants, or inflammatory MLE-12 cells (MLE-12 Inflam ), in the absence of additional external stimuli (A) . MCP-1 production in co-cultures of naïve MPI cells and MLE-12 Inflam , or MLE-12 Inflam treated with ICAM-1 blocking antibodies (B) . Cells were cultured together and cytokines were quantified after 24h via ELISA. Data are representative of three independent experiments and expressed as mean ± SD for three biological replicates. Multiple pairwise comparisons were performed by Prism software using One-Way ANOVA followed by Dunnett’s post hoc test. **p-value < 0.01; ***p-value < 0.001. ns was indicated when the differences were found to be non-significant.

Article Snippet: For ICAM-1 neutralization, MLE-12 monolayers were incubated with 5 μg/mL of ICAM-1 blocking antibody (Mouse ICAM-1/CD54, R&D Systems) for 1h prior to addition of MPI cells, as previously reported ( ).

Techniques: Blocking Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Software

Journal: Journal of Cell Communication and Signaling

Article Title: Stent treatment improves cerebral microcirculatory disorder and blood–brain barrier function in internal carotid artery stenosis via intercellular adhesion molecule 1 modulation

doi: 10.1002/ccs3.70058

Figure Lengend Snippet: Transcriptomic analysis of rat internal carotid artery severe stenosis model before and after stent implantation. (A) Experimental flowchart illustrating the collection of brain tissue samples and transcriptomic analysis process before and after stent implantation in rats; (B) differential gene expression analysis results, showing the gene expression differences between brain tissue samples before and after stent implantation; (C, D) clusterProfiler enrichment analysis results, displaying the enrichment of DEGs in gene ontology and Kyoto encyclopedia of genes and genomes pathways; (E) correlation analysis between ICAM1 and genes related to inflammation, BBB function, and cerebral microcirculation; (F) protein–protein interaction network analysis results, demonstrating the central role of ICAM1 in the network and its interactions with other inflammation‐related proteins; (G) gene set enrichment analysis enrichment analysis results, indicating significant enrichment of the NF‐κB signaling pathway in DEGs after stent implantation; (H, I) western blot results, showing the nuclear translocation of NF‐κB p65 and changes in the expression levels of ICAM1 and IκBα after stent implantation. All data are presented as mean ± standard error. Experiments were repeated three times, and statistical analysis was performed using ANOVA with Tukey post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. DEGs, differentially expressed genes.

Article Snippet: ELISA assays for TNF‐α (ab236712, Abcam, UK), IL‐6 (ab234570, Abcam, UK), MMP‐9 (R6000B, R&D Systems, USA), and ICAM‐1 (RIC100, R&D Systems, USA) were implemented as per the manufacturers' protocols.

Techniques: Gene Expression, Western Blot, Translocation Assay, Expressing

Journal: Journal of Cell Communication and Signaling

Article Title: Stent treatment improves cerebral microcirculatory disorder and blood–brain barrier function in internal carotid artery stenosis via intercellular adhesion molecule 1 modulation

doi: 10.1002/ccs3.70058

Figure Lengend Snippet: Protective effects of ICAM1 knockdown on HBMEC and SH‐SY5Y cells under OGD conditions. (A) Schematic diagram of the experimental procedure, showing the experimental design for ICAM1 knockdown and OGD treatment; (B) western blot analysis of ICAM1 expression in HBMECs; (C) detection of reactive oxygen species levels in HBMECs using DCF‐DA fluorescence probe; (D) time‐dependent changes in transendothelial electrical resistance (Ω▪cm 2 ) values of HBMECs over 90 min; (E) immunofluorescence staining of tight junction proteins (ZO‐1 and occludin) in HBMECs; (F) RT‐qPCR analysis of antioxidant‐related genes (Nrf2 and HO‐1) expression in HBMECs; (G) RT‐qPCR analysis of inflammatory cytokines (TNF‐α and IL‐6) expression in HBMECs. All data are presented as mean ± standard error of the mean. Experiments were repeated three times, and statistical analysis was performed using ANOVA followed by Tukey's post‐hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HBMEC, human brain microvascular endothelial cell; OGD, oxygen‐glucose deprivation; RT‐qPCR, reverse transcription quantitative polymerase chain reaction.

Article Snippet: ELISA assays for TNF‐α (ab236712, Abcam, UK), IL‐6 (ab234570, Abcam, UK), MMP‐9 (R6000B, R&D Systems, USA), and ICAM‐1 (RIC100, R&D Systems, USA) were implemented as per the manufacturers' protocols.

Techniques: Knockdown, Western Blot, Expressing, Fluorescence, Immunofluorescence, Staining, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Cell Communication and Signaling

Article Title: Stent treatment improves cerebral microcirculatory disorder and blood–brain barrier function in internal carotid artery stenosis via intercellular adhesion molecule 1 modulation

doi: 10.1002/ccs3.70058

Figure Lengend Snippet: Effect of ICAM1 knockdown on HBMEC function. (A) Schematic diagram of the experimental procedure, illustrating the experimental design for ICAM1 knockdown and oxygen–glucose deprivation treatment in HBMEC and SH‐SY5Y cells; (B) CCK‐8 assay to assess HBMEC cell proliferation; (C) Transwell migration assay to evaluate HBMEC cell migration capacity; (D) scratch assay to measure the migration rate of HBMEC cells; (E) tube formation assay to assess the in vitro angiogenic ability of HBMECs; (F) annexin V‐FITC/PI flow cytometry to determine the apoptosis rate of HBMEC cells. Data are presented as mean ± standard error of the mean. All experiments were performed in triplicate, and statistical analysis was conducted using ANOVA followed by Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HBMEC, human brain microvascular endothelial cell.

Article Snippet: ELISA assays for TNF‐α (ab236712, Abcam, UK), IL‐6 (ab234570, Abcam, UK), MMP‐9 (R6000B, R&D Systems, USA), and ICAM‐1 (RIC100, R&D Systems, USA) were implemented as per the manufacturers' protocols.

Techniques: Knockdown, CCK-8 Assay, Transwell Migration Assay, Migration, Wound Healing Assay, Tube Formation Assay, In Vitro, Flow Cytometry

Journal: Journal of Cell Communication and Signaling

Article Title: Stent treatment improves cerebral microcirculatory disorder and blood–brain barrier function in internal carotid artery stenosis via intercellular adhesion molecule 1 modulation

doi: 10.1002/ccs3.70058

Figure Lengend Snippet: Effects of stent implantation on cerebral microcirculation, cognitive function, and BBB in rats with severe internal carotid artery stenosis. (A) Diagram illustrating experimental groups and protocol; (B) laser speckle contrast imaging measurement of cerebral blood flow in rats; (C) Morris water maze test to assess spatial learning and memory in rats; (D) balance beam test to evaluate motor coordination in rats; (E) H&E staining to observe pathological changes in rat brain tissue structure; (F) western blot analysis of the expression levels of BBB tight junction proteins occludin and ZO‐1 in rat brain tissue; (G, H) ELISA measurement of inflammatory cytokines (TNF‐α, IL‐6) and the expression levels of ICAM1 and MMP‐9 in rat brain tissue. Data are presented as mean ± standard error of the mean, with experiments repeated three times and 10 rats per group. Statistical analysis was performed using ANOVA followed by Tukey's post‐hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: ELISA assays for TNF‐α (ab236712, Abcam, UK), IL‐6 (ab234570, Abcam, UK), MMP‐9 (R6000B, R&D Systems, USA), and ICAM‐1 (RIC100, R&D Systems, USA) were implemented as per the manufacturers' protocols.

Techniques: Imaging, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Nutrition

Article Title: In vivo effects of javamide-I/-II on metabolic, hepatic, cardiovascular and inflammatory risk factors

doi: 10.3389/fnut.2025.1661468

Figure Lengend Snippet: Effects of J1/J2 on sICAM and sE-selectin. sICAM (A) and sE-selectin (B) were measured using plasma samples. There were no significant differences in sICAM and sE-selectin levels between the CG and JG groups. Data is presented as the mean ± SD ( n = 10).

Article Snippet: Rat soluble intercellular adhesion molecule 1 (sICAM) level was determined in plasma samples using sICAM ELISA kit (Cat # RIC100, R&D systems, Minneapolis, MN, the United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics