Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Bone Marrow Mesenchymal Stem Cell-Derived Dermcidin-Containing Migrasomes enhance LC3-Associated Phagocytosis of Pulmonary Macrophages and Protect against Post-Stroke Pneumonia.

doi: 10.1002/advs.202206432

Figure Lengend Snippet: Figure 7. DCD is beneficial to AIS recovery and DCD-containing BM-MSC-derived migrasome effectively promotes phagocytosis of macrophages. A– G) Peripheral blood of AIS patients (acute phase, 0–3d after disease onset, n = 16) and healthy controls (HC, n = 8) were collected. (A) Plasma DCD concentration was assessed with ELISA. *p < 0.05, compared with HC by Student’s t-test (mean ± standard deviation). (B) Correlation of clinic parameters and plasma DCD concentration was assessed with Spearman correlation analysis and Point-biserial correlations. *p < 0.05. DM, diabetes mellitus, CHD, coronary heart disease. (C) Representative images of the magnetic resonance diffusion weighted imaging (MR-DWI) of AIS patients with low plasma DCD concentration (DCD ≤3.33 ng ml−1) or high plasma DCD concentration (DCD > 3.33 ng ml−1). (D) Association between plasma DCD concentration with infarct scale was estimated with Spearman correlation analysis. (E) Association between plasma DCD concentration with delta NIHSS (NIHSS at 7d minus NIHSS at 1d) was estimated with Spearman correlation analysis. (F) Representative images of the chest Computed Tomography (CT) of AIS patients with low plasma DCD concentration (DCD ≤3.39 ng ml−1, median of the cohort) or high plasma DCD concentration (DCD > 3.39 ng ml−1, median of the cohort). (G) Pie charts showing the occurrence of post-stroke pneumonia in AIS patients with low and high plasma DCD concentrations. H) DCD (1 ng ml−1), PBS-migrasomes (PBS-M, 50 μg ml−1) or E. Coli-migrasomes (E. Coli-M, 50 μg ml−1) labeled with Dil (red) were treated to BMDM (15 min). Immunostaining of WGA (green) and DCD (withe) in migrasome-treated BMDM was performed. Experiments were repeated for three times. I,J) BMDM were first pre-stimulated with DCD (1 ng ml−1), PBS-M (50 μg ml−1), or E. Coli-M (50 μg ml−1) for overnight then treated with E. Coli (E. Coli : BMDM = 20:1, 1 h). Phagocytic efficiency of BMDM to GFP expressing E. Coli was assessed with flow cytometry (I) and immunostaining (J). Experiments were repeated three times. **p < 0.01, compared with PBS-treated group by one-way ANOVA (mean ± standard deviation).

Article Snippet: Human Monocyte Enrichment and Macrophage Differentiation: Mononucleus cells were isolated from peripheral blood of healthy adults (age = 18–40y) with human peripheral blood monocyte isolation Solution kit (Solarbio, P8680).

Techniques: Derivative Assay, Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Imaging, Computed Tomography, Labeling, Immunostaining, Expressing, Cytometry

Journal: Science Advances

Article Title: Endothelial CLEC5A drives barrier dysfunction and vascular leakage responsible for lung injury in bacterial pneumonia and sepsis

doi: 10.1126/sciadv.adt7589

Figure Lengend Snippet: ( A ) UMAP plot of monocyte clusters in WT lungs ( n = 3) and CLEC5A −/− lungs ( n = 3) and percentage in immune cells. ( B ) Dot plot showing the expression of adhesion molecules in each subtype of endothelial cells. ( C ) GSEA of the DEGs in the endothelial cells between CLEC5A −/− and WT lungs showing significant enrichment of cell-adhesion related-GO pathways, in which the up-regulated DEGs were mainly enriched. The significance was determined by P value with false discovery rate (FDR) < 0.25. ( D ) Mouse PMVECs were isolated and challenged by LPS (10 μg/ml) for 24 hours. ( E ) Levels of MCP-1 and VCAM-1 in the supernatant by ELISA. ( F ) Relative mRNA expression of MCP-1 and VCAM-1 in PMVECs. ( G ) Overview of in vitro adhesion and trans-endothelial migration assays in mouse PMVECs and HUVECs. ( H ) Mouse PBMCs were labeled by BCECF-AM, and the adherent monocytes to PMVECs were observed under a fluorescence microscopy. Scale bar, 50 μm. Monocyte-endothelial adhesion was quantified as the percentage of fluorescence intensity relative to the WT group. ( I ) Transmigration of mouse PBMCs across the PMVEC monolayer was shown as the percentage of transmigrated cells relative to the WT group by MTT assay. Lentivirus-mediated gene delivery for CLEC5A (CLEC5A oe ) or shRNA targeting CLEC5A (CLEC5A sh ) were carried out in HUVECs. ( J ) Adhesion of THP-1 to HUVECs was shown as fluorescence and quantified as the percentage of fluorescence intensity relative to the control group. Scale bar, 50 μm. ( K ) Migration of THP-1 through the HUVEC monolayer shown as the percentage of transmigrated cells relative to the control group. The statistical significance was determined by an unpaired two-tailed t test (or with Welch’s correction) for comparison between two groups and by one-way ANOVA test for comparisons among four groups (biological replicates, n = 4 per group for each experiment).

Article Snippet: Mouse peripheral blood monocytes (PBMCs) were isolated and purified using the commercial kit (P5230#, Solarbio, Beijing, China).

Techniques: Expressing, Isolation, Enzyme-linked Immunosorbent Assay, In Vitro, Migration, Labeling, Fluorescence, Microscopy, Transmigration Assay, MTT Assay, shRNA, Control, Two Tailed Test, Comparison