Journal: BMC Neuroscience

Article Title: Exploring extracellular vesicle surface markers and microRNA regulation following remote ischemic conditioning in patients with stroke; a randomized-controlled pilot study

doi: 10.1186/s12868-025-00993-1

Figure Lengend Snippet: EV surface marker changes after RIC treatment: The expression of the EV-markers CD62 and MCP-1 is plotted showing the Log 2 fold changes compared to the baseline sample of each patient. The p-value is shown for both the significant changes within RIC group (red) and compared with the miRNA expression changes over time in Sham group (grey). ( a ) CD62 shows an upregulation in RIC treated patients compared to Sham. ( b ) MCP-1 showed no significant changes in between the different timepoints

Article Snippet: CD62 E/P , R & D Systems , BBA1 , BBIG-E(13D5).

Techniques: Marker, Expressing

Journal: BMC Neuroscience

Article Title: Exploring extracellular vesicle surface markers and microRNA regulation following remote ischemic conditioning in patients with stroke; a randomized-controlled pilot study

doi: 10.1186/s12868-025-00993-1

Figure Lengend Snippet: Correlation analysis of significant miRNA measurements (hsa-miR-19b-3p, hsa-miR-30d-5p, hsa-miR-374a-5p, hsa-miR-20a/20b-5p, hsa-miR-24-3p) and CD62/P-selectin. The histograms on the diagonal show the distribution of each variable. The lower triangular panels depict scatter plots with fitted lines, illustrating the linear relationships between pairs of variables. The upper triangular panels display the correlation coefficients, with significance levels indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: CD62 E/P , R & D Systems , BBA1 , BBIG-E(13D5).

Techniques:

Journal: medRxiv

Article Title: Endothelial cell-activating antibodies in COVID-19

doi: 10.1101/2021.01.18.21250041

Figure Lengend Snippet: A , Schematic workflow for in-cell ELISA. HUVEC were cultured for 6 hours with serum from either healthy controls (collected pre-pandemic) (n=38) or patients hospitalized with COVID-19 (n=118). Cells were then fixed and surface expression of E-selectin ( B ), VCAM-1 ( C ), or ICAM-1 ( D ) was quantified. Median values are indicated by horizontal lines. Groups were analyzed by Mann-Whitney test; ****p<0.0001. E , Beyond the 118 COVID-19 patients tested in panel D, surface expression of ICAM-1 was tested in the context of an additional 126 unique patient samples. Patients requiring mechanical ventilation (n=101) are compared to hospitalized patients who were not mechanically ventilated (n=143); p<0.01 by Mann Whitney test. F , Serum from healthy controls (n=38) and COVID-19 patients (n=102) were assessed for soluble E-selectin. COVID-19 samples were compared to controls by Mann-Whitney test; ****p<0.0001. G , Soluble E-selectin was compared to HUVEC E-selectin expression as presented in panel B. Correlation was determined by Spearman’s method. H , HUVEC were cultured for 6 hours with plasma from healthy controls (n=36), intensive care unit patients with non-COVID sepsis (n=100), or patients hospitalized with COVID-19 (n=72). Cells were then fixed and surface expression of ICAM-1 was quantified. Groups were analyzed by one-way ANOVA with correction for multiple comparisons by Holm-Sidak’s method; *p<0.05 and ***p<0.001.

Article Snippet: After washing with PBS, cells were incubated with 5 µg/ml primary mouse anti-human antibodies against E-selectin (catalog BBA26, R&D), VCAM-1 (catalog BBA5, R&D), or ICAM-1 (ab2213, Abcam) at 4°C overnight.

Techniques: In-Cell ELISA, Cell Culture, Expressing, MANN-WHITNEY

Journal: medRxiv

Article Title: Endothelial cell-activating antibodies in COVID-19

doi: 10.1101/2021.01.18.21250041

Figure Lengend Snippet: HUVEC were cultured for 6 hours with serum from either healthy controls (collected pre-pandemic) (n=38) or patients hospitalized with COVID-19 (n=118). Cells were then fixed and surface expression of E-selectin ( A ), VCAM-1 ( B ), or ICAM-1 ( C ) was quantified. Median values are indicated by horizontal lines. Data were presented as raw absorbance (optical density 650 nm). Groups were analyzed by Mann-Whitney test; ****p<0.0001.

Article Snippet: After washing with PBS, cells were incubated with 5 µg/ml primary mouse anti-human antibodies against E-selectin (catalog BBA26, R&D), VCAM-1 (catalog BBA5, R&D), or ICAM-1 (ab2213, Abcam) at 4°C overnight.

Techniques: Cell Culture, Expressing, MANN-WHITNEY

Journal: medRxiv

Article Title: Endothelial cell-activating antibodies in COVID-19

doi: 10.1101/2021.01.18.21250041

Figure Lengend Snippet: Soluble E-selectin in COVID-19 serum was compared to laboratory and clinical data when available on the same day as the serum collection. Spearman’s correlations are presented for C-reactive protein (n=83) ( A ), D-dimer (n=72) ( B ), calprotectin (n=102) ( C ), oxygenation efficiency (n=99) (pulse oximetry/fraction of inspired oxygen, D ).

Article Snippet: After washing with PBS, cells were incubated with 5 µg/ml primary mouse anti-human antibodies against E-selectin (catalog BBA26, R&D), VCAM-1 (catalog BBA5, R&D), or ICAM-1 (ab2213, Abcam) at 4°C overnight.

Techniques:

Journal: medRxiv

Article Title: Endothelial cell-activating antibodies in COVID-19

doi: 10.1101/2021.01.18.21250041

Figure Lengend Snippet: A , Serum was pooled from 3 patients with positive aCL IgG or 5 patients with positive aPS/PT IgG. HUVEC monolayers were then treated with 2.5% COVID or control serum for 6 hours. Calcein-AM-labeled neutrophils were then added as described in Methods. Scale bar=200 microns. Mean ± standard deviation is presented for n=3 independent experiments; *p<0.05 by one-way ANOVA corrected by Dunnett’s test. B-D , IgG was depleted from each of the aforementioned pools. Activation of HUVEC was determined after culture for 6 hours as defined by surface expression of E-selectin ( B ), VCAM-1 ( C ), or ICAM-1 ( D ). The experiment was repeated on 3 different days, and bars represent mean and standard deviation. Groups were compared by 2-sided paired t-test; *p<0.05 and **p<0.01. E , IgG was purified from the pooled samples referenced in A-C, and then supplemented (100 μg/ml) into control serum that had been depleted of IgG. Activation of HUVEC was determined after culture for 6 hours as defined by surface expression of ICAM-1. Groups were compared by one-way ANOVA with correction for multiple comparisons by Tukey’s test; **p<0.01.

Article Snippet: After washing with PBS, cells were incubated with 5 µg/ml primary mouse anti-human antibodies against E-selectin (catalog BBA26, R&D), VCAM-1 (catalog BBA5, R&D), or ICAM-1 (ab2213, Abcam) at 4°C overnight.

Techniques: Labeling, Standard Deviation, Activation Assay, Expressing, Purification

Journal: Frontiers in Immunology

Article Title: Molecular determinants of STEC-HUS: from complement activation to microvascular thrombosis

doi: 10.3389/fimmu.2026.1749811

Figure Lengend Snippet: Prothrombotic effects of STEC-HUS serum are dependent on WPB exocytosis from endothelial cells. P-selectin (A) and vWF (B) expression on unstimulated HMEC-1 exposed to serum from patients with acute STEC-HUS (n = 5 for P-selectin experiments; n = 6 for vWF experiments), in the presence or in the absence of different complement inhibitors (sCR1, 150 µg/mL; factor B inhibitor, iptacopan,10 µM; eculizumab 100 µg/mL) or to a pool of control sera (normal human serum, NHS), run in parallel. Results are shown as pixel 2 /high-power field (HPF) of stained surface area. Data are mean ± SD. Circles represent single patients’ data. The addition of either sCR1, or iptacopan or eculizumab to patient’s serum significantly decrease both P-selectin and vWF expression induced by patient’s serum alone. *P < 0.05 vs STEC-HUS serum alone (ANOVA, followed by Tukey’s multiple comparisons test for data of P-selectin expression and by Holm-Šídák’s multiple comparisons test for data of vWF expression). (C) Endothelial surface area covered by thrombi on ADP-activated HMEC-1 exposed to serum from STEC-HUS patients collected during the acute phase of the disease and then perfused with whole blood. Before the experiments, HMEC-1 were left for 16 hours with medium added or not with the RalA inhibitor BQU57 (10 µM). Data are expressed as mean ± SD of percentages of serum-induced thrombus formation in respect to a pool of control sera (normal human serum, NHS), run in parallel in each experiment and set as 100% (n = 3 independent experiments). Circles indicate single patients’ data. Horizontal dashed lines indicate upper and lower limits of the normal range . *P < 0.05 (paired Student’s t test). (D) Representative confocal microscopy images (original magnification X200) of experiments of thrombus formation (green staining) relative to Figure 6C . Scale bar: 100 µm.

Article Snippet: Cells were washed again and treated with the following specific antibodies: FITC-conjugated rabbit anti-human C3c-complement (Dako, that recognizes C3c, part of C3 and C3b, 1:300 final dilution in Dapi 1 μg/mL); or rabbit anti-human complement C5b-9 complex (Calbiochem, 1:200 final dilution in PBS1X) followed by FITC-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:50 final dilution in 1 μg/mL Dapi); or goat anti-human C4 (Abcam, 1:100 final dilution in PBS1X) followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:200 final dilution in Dapi 1 μg/mL); or FITC-conjugated anti-human IgG (Sigma Aldrich, 1:32 final dilution in 1 μg/mL Dapi); or mouse anti-human P-selectin (R&D System, 20 μg/mL final concentration in PBS1X), followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:60 final dilution in 1 μg/mL Dapi); or rabbit anti-human vWF (Dako, 10 μg/mL final concentration in PBS1X), followed by Cy3-conjugated secondary antibody (Jackson ImmunoResearch Laboratories, 1:50 final dilution in 1 μg/mL Dapi).

Techniques: Expressing, Control, Staining, Confocal Microscopy