Journal: Cell Reports Medicine

Article Title: Artificial exosomes synergistically reshape sepsis immune homeostasis by modulating neutrophil fate and blocking PD-1/PD-L1

doi: 10.1016/j.xcrm.2026.102819

Figure Lengend Snippet: Preparation, characterization, and interaction of AT@NV-PD1 nanoparticles with neutrophils (A) CLSM images show PD-1 (green) expression on engineered RAW 264.7 membranes (scale bars, 15 μm). (B and C) Flow cytometry quantifies PD-1 expression in primary vs. engineered cells. (D) Western blot analysis of PD-1 expression in primary and genetically engineered RAW 264.7 cells. (E and F) DLS and TEM reveal AT@NV-PD1’s hydrodynamic diameter (110–130 nm) and core-shell structure (scale bars, 50 nm). (G) CLSM demonstrates AT@NV-PD1 (red) uptake by activated neutrophils (scale bars, 10 μm). (H) Anti-PD-L1 pretreatment blocks uptake (scale bars, 10 μm). (I) Resting neutrophils show minimal uptake (scale bars, 10 μm). (J) Flow cytometry shows elevated PD-L1 on septic neutrophils. (K and L) Enhanced AT@NV-PD1 uptake in CLP-model neutrophils vs. healthy controls. (M–Q) Nanodecoys effectively neutralize inflammatory mediators (TNF-α, IL-1β, IL-6, PD-L1, LPS). All data are presented as mean ± SD ( n = 3), ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, ns, not significant.

Article Snippet: Lipopolysaccharide (LPS) , MedChemExpress (USA) , Cat. HY-D1056A1.

Techniques: Expressing, Flow Cytometry, Western Blot

Journal: Cell Reports Medicine

Article Title: Artificial exosomes synergistically reshape sepsis immune homeostasis by modulating neutrophil fate and blocking PD-1/PD-L1

doi: 10.1016/j.xcrm.2026.102819

Figure Lengend Snippet: AT@NV-PD1 regulates neutrophil fate and its mechanism in vitro (A and B) Flow cytometry analysis and quantitative results showing that AT@NV-PD1 treatment can significantly induce apoptosis in neutrophils stimulated by IFN-γ and LPS. (C) Representative immunofluorescence images of NETs; CitH3 (red), MPO (green), and DAPI (blue) staining, (scale bars, 20 μm). (D) Representative CLSM images of SYTOX Green-stained NETs. Nuclear DNA was stained with DAPI (blue), and extracellular DNA was stained with SYTOX Green (green), (scale bars, 20 μm). (E and F) Expression levels of NETs and IL-6 in neutrophils treated with different formulations, as measured by ELISA. Data are presented as mean ± standard deviation. Differences were evaluated by one-way ANOVA followed by Tukey’s multiple comparisons test, n = 5, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗ p < 0.0001.

Article Snippet: Lipopolysaccharide (LPS) , MedChemExpress (USA) , Cat. HY-D1056A1.

Techniques: In Vitro, Flow Cytometry, Immunofluorescence, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Cell Reports Medicine

Article Title: Artificial exosomes synergistically reshape sepsis immune homeostasis by modulating neutrophil fate and blocking PD-1/PD-L1

doi: 10.1016/j.xcrm.2026.102819

Figure Lengend Snippet: AT@NV-PD1 attenuates T cell exhaustion and promotes immune function recovery and does not impair neutrophil generation in the bone marrow (A) Flow cytometry analysis of the CD4 + /CD8 + T cell ratio in peripheral blood from CLP mice 5 days after intervention with different formulations. (B and C) Representative flow cytometry plots (B) and quantitative analysis (C) of CD4 + T cells in peripheral blood. (D and E) Representative images (D) and quantification (E) of TUNEL staining for apoptosis in splenocytes from healthy control and sepsis-treated mice (scale bars, 100 μm). (F and G) Representative flow cytometry plots (F) and quantitative analysis (G) of Treg cells in peripheral blood. (H) Flow cytometry plots of CD11b + cells in peripheral blood. (I and J) Representative flow cytometry plots and quantitative analysis of M-MDSC cells in peripheral blood. (K) Mouse body weights were monitored during the experiment, including healthy mice, PBS-treated sepsis mice, and AT@NV-PD1 -treated sepsis mice (AT7519 at a dose of 5 mg/kg). (L–O) Mice were subjected to intraperitoneal (i.p.) LPS injection to establish a sepsis model, and AT@NV-PD1 treatment was administered 4 h after LPS administration. Four hours later, LPS-challenged mice were intravenously (i.v.) injected with AT@NV-PD1 (AT7519 at a dose of 5 mg/kg). Control mice received neither LPS nor AT@NV-PD1 treatment. After 72 h, all surviving mice and control (healthy) mice were challenged with LPS (i.t. [intratracheally], 10 mg/kg). At 84 h, bronchoalveolar lavage fluid (BALF) was collected to assess neutrophil counts, IL-1β, TNF-α, and IL-6 levels. All data are presented as mean ± standard deviation (SD), and differences were evaluated by one-way ANOVA followed by Tukey’s multiple comparisons test ( n = 5; ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001).

Article Snippet: Lipopolysaccharide (LPS) , MedChemExpress (USA) , Cat. HY-D1056A1.

Techniques: Flow Cytometry, TUNEL Assay, Staining, Control, Injection, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: Targeting IgG Autoantibodies for Improved Cytotoxicity of Bactericidal Permeability Increasing Protein in Cystic Fibrosis

doi: 10.3389/fphar.2020.01098

Figure Lengend Snippet: Elevated levels of BPI present in plasma and BAL of people with CF. (A , B) LPS levels in CF plasma and BAL correlated with CF-ABLE score. Quantification of an association between variables was achieved by Spearman correlation. (C , D) Comparative analysis of BPI present in plasma or BAL of Phe508del homozygous PWCF (CF), healthy controls (HC) or NCFB patients was performed by ELISA. (C) BPI levels were significantly increased in plasma of CF compared to HC (n=22 and n=14 subjects per group, respectively, p=0.007, Mann Whitney U-test). (D) BAL levels of BPI were significantly increased in CF (n=5) compared to HC (n=4) or NCFB patients (n=5) (p<0.0001, One-way ANOVA, followed by Bonferroni post-hoc test for selected groups). (E) BAL samples from HC, CF, NCFB or COPD were subjected to SDS-PAGE and Western blot analysis for BPI. An immuno-band of increased intensity for BPI was detected in CF BAL samples (top panels). Lower panel, a control immunoblot to ensure BPI specificity. The blot was halved, with one half probed with secondary antibody only with no BPI immune-bands visible (↑ indicates where blot was cut). Human BPI (hBPI) was used as a positive control. All measurements are means ± SEM from biological replicates.

Article Snippet: Lipopolysaccharide (LPS) was quantified in plasma and BAL of PWCF ( ) using an LPS ELISA (Cusabio: catalog number CSB-E09945h).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, SDS Page, Western Blot, Control, Positive Control

Journal: Frontiers in Pharmacology

Article Title: Targeting IgG Autoantibodies for Improved Cytotoxicity of Bactericidal Permeability Increasing Protein in Cystic Fibrosis

doi: 10.3389/fphar.2020.01098

Figure Lengend Snippet: Increased levels of plasma autoantibodies and IgG-bound BPI in CF airway samples. (A) IgG autoantibodies against BPI were quantified in plasma of PWCF (CF, n=30) or healthy controls (HC, n=37). A significant increase in the titre of BPI autoantibodies were detected in CF (p<0.0001, Mann-Whitney U test). Positivity was set as 10 U/ml as indicated by the hatched line. All CF samples were positive for BPI autoantibodies. Increased circulating IgG BPI antibody level in CF individuals determined by ELISA. (B) Plasma samples from HC (n=38), CF (n=28) and CF individuals receiving ivacaftor treatment (n=10). No difference in levels of circulating anti-BPI IgG autoantibodies between CF and CF individuals receiving ivacaftor treatment (p=0.39). (C) Representative Coomassie blue stained SDS gel of purified IgG from CF BAL. Protein G Sepharose was used to isolate IgG from BAL of Phe508del homozygous PWCF. Starting BAL sample (St), unbound material (Un) or purified bound IgG (Bd) with or without DTT reduction are presented. Purified IgG (150 kDa, closed arrow), and the heavy (50 kDa) and light chains (25 kDa) of IgG are indicated (open arrows) (1 representative images of n = 5 biological repeats). (D) Protein G Sepharose was used to isolate IgG-BPI complexes from CF BAL. Reactions were analyzed by immunoblotting for BPI positivity using a mouse monoclonal anti-BPI antibody. Starting BAL sample (St), unbound material (Un) or IgG-bound BPI (Bd) are presented. Human BPI (hBPI) was used as a positive control (55 kDa). A control immunoblot to ensure BPI specificity omitted primary antibody (right hand panel), with no immune-band apparent. 3 representative images of n=5 biological repeats.

Article Snippet: Lipopolysaccharide (LPS) was quantified in plasma and BAL of PWCF ( ) using an LPS ELISA (Cusabio: catalog number CSB-E09945h).

Techniques: Clinical Proteomics, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Staining, SDS-Gel, Purification, Western Blot, Positive Control, Control

Journal: International Journal of Molecular Medicine

Article Title: Remimazolam alleviates myocardial ischemia/reperfusion injury and inflammation via inhibition of the NLRP3/IL-1β pathway in mice

doi: 10.3892/ijmm.2025.5498

Figure Lengend Snippet: Rema alleviates LPS-induced release and increased expression levels of IL-1β, IL-6 and TNF-α in cultured Raw264.7 cells. (A) Experimental protocol using cultured Raw264.7 macrophages. (B) LPS induced alterations in the release of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema inhibited these LPS-mediated changes (n=3). (C) LPS induced alterations in the gene expression levels of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema inhibited these effects (n=3). (D) DAPA inhibited the LPS-induced release of IL-1β, IL-6 and TNF-α in Raw264.7 cells, while treatment with Rema did not affect the alterations induced by DAPA (n=3). *** P<0.001. DAPA, dapansutrile; LPS, lipopolysaccharide; ns, not significant; Rema, remimazolam; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Cells were cultured in DMEM without FBS and pre-treated with 100 μ g/ml Rema (diluted in DMEM) for 20 min, and subsequently treated with lipopolysaccharide (LPS; 0.5 μ g/ml diluted in DMEM; cat. no. HY-D1056H, MedChemExpress) for 24 h in incubator at 37°C.

Techniques: Expressing, Cell Culture, Gene Expression, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction