Journal: ASAIO journal (American Society for Artificial Internal Organs : 1992)

Article Title: Benchtop von Willebrand Factor Testing: Comparison of Commercially Available VADs and Evaluation of Variables for a Standardized Test Method

doi: 10.1097/MAT.0000000000000849

Figure Lengend Snippet: Normal FFP, FFP+rhADAMTS13 (1 or 4μg/mL) and FFP+EDTA (10mM) were assayed for ADAMTS13 antigen level(A) and activity(B). The double solid line (=) denotes normal levels. Error bars are +/− one standard deviation.

Article Snippet: Aliquots supplemented with 1 or 4μg/mL recombinant human ADAMTS13 (rhADAMTS13, R&D Systems, Minneapolis, MN), or 10mM ethylenediaminetetraacetic acid (EDTA) were rocked on a VariMix platform rocker (Model {"type":"entrez-nucleotide","attrs":{"text":"M79735","term_id":"271754","term_text":"M79735"}} M79735 , Thermo Scientific) at 1Hz, with samples collected into low protein retention microcentrifuge (LPRM) tubes (Fisher Scientific, Pittsburgh, PA) at 30 and 60 minutes, then hourly until 6h.

Techniques: Activity Assay, Standard Deviation

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 1. Mean (±SEM) microsphere- derived risk area from animals assigned to either (A) 30 min or (B) 45 min of coronary occlusion. C, Mean (±SEM) microvascular blood flow from myocardial contrast echocardiography (MCE) performed during coronary occlusion in the risk area and remote territories, as well as from sham-treated mice. D, Example of background-subtracted, color-coded MCE images during coronary occlusion, and time-intensity data after a destructive pulse sequence from the risk area and remote territory. For images, time after destructive pulse is at upper left and color-coded scale at bottom of the 5 s image. ADAMTS13 indicates a disintegrin and metalloprotein- ase with a thrombospondin type-1 motif member 13; LV, left ventricle; RA, risk area; and WT, wild-type. *P <0.05 vs remote territory.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Derivative Assay, Sequencing

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 2. Postischemic myocardial perfusion by MCE in mice undergo- ing 30 min of ischemia and reperfusion measured in the remote region and the risk area. Data (mean±SEM) are shown for (A) myocardial microvascular blood flow (MBF), (B) microvascular blood volume (MBV), and (C) microvascular flux rate (β). ADAMTS13 indicates a disintegrin and metalloproteinase with a thrombo- spondin type-1 motif member 13. *P <0.05 vs remote territory; †P <0.05 vs WT (wild type); ‡P <0.05 only before correction for multiple comparison.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Comparison

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 3. Mean (±SEM) signal enhance- ment on MCE molecular imaging after 30 min of ischemia and reperfusion for (A) VWF (von Willebrand factor) A-1 domain and (B) platelet GP (glycoprotein) Ibɑ. Data are expressed as signal difference between targeted and control microbubbles (MB). C, Examples of color-coded MCE molecular imaging in the short-axis plane for targeted and control MBs (color scale at bottom) and the microsphere-derived risk area in the same short-axis plane. *P <0.05 vs remote territory; †P <0.05 vs wild-type and ADAMTS13−/− (a disintegrin and metal- loproteinase with a thrombospondin type-1 motif member 13); ‡P <0.05 vs WT (wild type).

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Imaging, Control, Derivative Assay

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 5. Mean (±SEM) infarct size by triphenyltetrazolium chloride staining as a percentage of the total microsphere-de- rived risk area for mice undergoing either (A) 30 min of ischemia followed by 90 min of reperfusion or (B) 45 min of ischemia followed by 3 d of reperfusion. *P <0.05 vs WT (wild type) before Bonfer- roni correction; †P <0.05 vs WT+ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type-1 motif member 13) after Bonferroni correction.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Staining

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 4. Examples of immunohistochemis- try from the sham-treated control WT (wild type) mice (anterior and posterior myocar- dial with anterior labeled as risk area) and from postischemic WT and ADAMTS13−/− (a disintegrin and metalloproteinase with a thrombospondin type-1 motif member 13) mice. The top and bottom rows show fused im- ages for platelet CD41 immunostaining (red), endothelial lectin staining (green), and nuclear counterstain (blue). The middle row shows corresponding images for only the red channel (platelet CD41) in the risk area. For the insets, arrowheads show single platelets, whereas the arrow in the ADAMTS13−/− mouse illustrates 3 platelets linearly arranged (scale bar=20 μm). MI indicates myocardial infarction.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Control, Labeling, Immunostaining, Staining

Journal: medRxiv

Article Title: Distinct impact of IgG subclass and Fc-FcγR interaction on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.1101/2020.12.07.20243774

Figure Lengend Snippet: IgG4 is less pathogenic than IgG1 in anti-ADAMTS13 autoantibodies. (A) Schematic diagram of the experimental design. In brief, wide-type C57BL/6 or hFCGR Tg mice were treated with 10 μg of control human IgG (Ctrl hIgG, n ≥ 3), or anti-ADAMTS13 TTP1-420(IgG1) or TTP1-420(IgG4) (n ≥ 4) on day 0 through tail vein injection, blood was collected on day -1, day 1, day 5 and day 8 and analyzed for ADAMTS13 activity. (B-E) Plots showing ADAMTS13 activity in the plasma of WT (B, C) or hFCGR Tg (D, E) mice treated with the indicated antibodies at the indicated time points and analyzed by the FRETS-VWF73 assay, represented as relative fluorescence units (RFU) changing rates over time (RFU/min) (B, D), and the RFU change within 1 hour on day -1 and day 8 (C, E). Mean ± SEM values are plotted. Two-way ANOVA with Tukey’s (B, D) or Sidak’s (C, E) multiple comparisons tests. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. A representative of two independent experiments is shown.

Article Snippet: ADAMTS13 antigen levels in the plasma of HC and TTP patients were measured using Human ADAMTS13 Quantikine ELISA Kit (R&D Systems) according to the manufacturer’s instructions with minor modifications: (1) diluted plasma samples of HC (1:50) and TTP patients (1:2) were used; (2) the range of the standard curve was broadened to 0.78125-100 ng/ml.

Techniques: Injection, Activity Assay, Fluorescence

Journal: medRxiv

Article Title: Distinct impact of IgG subclass and Fc-FcγR interaction on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.1101/2020.12.07.20243774

Figure Lengend Snippet: The protective effect of the IgG4 subclass in anti-ADAMTS13 autoantibodies is due to reduced FcγR-mediated antibody effector function. (A) Schematic diagram of the experimental design. In brief, WT, FcγRα null , or Fcer1g -/- mice were treated and analyzed as in . (B-E) Plots showing ADAMTS13 activity in the plasma of WT and FcγRα null mice (B, C), WT and Fcer1g -/- mice (D, E), or WT mice (F, G) treated with the indicated antibodies and analyzed at indicated time points as in and , represented as RFU changing rates over time (RFU/min) (B, D, F), and the RFU change within 1 hour on the indicated days (C, E, G). Mean ± SEM values are plotted. Two-way ANOVA with Sidak’s (B-E and G) or Tukey’s (F) multiple comparisons tests. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. A representative of two independent experiments is shown.

Article Snippet: ADAMTS13 antigen levels in the plasma of HC and TTP patients were measured using Human ADAMTS13 Quantikine ELISA Kit (R&D Systems) according to the manufacturer’s instructions with minor modifications: (1) diluted plasma samples of HC (1:50) and TTP patients (1:2) were used; (2) the range of the standard curve was broadened to 0.78125-100 ng/ml.

Techniques: Activity Assay

Journal: medRxiv

Article Title: Distinct impact of IgG subclass and Fc-FcγR interaction on autoantibody pathogenicity in different IgG4-mediated diseases

doi: 10.1101/2020.12.07.20243774

Figure Lengend Snippet: ADAMTS13-specific IgG1 levels in the plasma of acquired TTP patients inversely correlate to the ADAMTS13 Ag levels and activity. (A-C) Plots showing ADAMTS13-specific IgG (A), ADAMTS13 activity (B), and ADAMTS13 antigen (C) levels in the plasma of HC (n = 23) and acquired TTP patients during the acute phase (n = 44). (D) Plot showing correlation analysis of ADAMTS13 activity with ADAMTS13 Ag concentration in TTP patients. (E) ADAMTS13-specific IgG1 (left panel) and IgG4 (right panel) levels in the plasma of TTP patients and HC. (F) Plot showing correlation analysis of ADAMTS13-specific IgG1 with IgG4 levels in TTP patients, with the threshold for IgG1 and IgG4 autoantibodies (two times of HC average values) annotated. (G-J) Plots showing correlation analysis between the anti-ADAMTS13 IgG1 (G, H) and IgG4 (I, J) levels with ADAMTS13 activity (G, I) and ADAMTS13 Ag (H, J) levels in TTP patients, respectively. (K) Plots showing IgG1 and IgG4 anti-ADAMTS13 levels in TTP plasma samples normalized to HC, with IgG1 dominant TTP and IgG4 dominant TTP samples annotated. (L, M) Plotting showing ADAMTS13 activity (L) and antigen (M) levels in TTP plasma samples as annotated in (K). Each symbol is derived from an individual plasma sample. Mean ± SEM values are plotted. Unpaired nonparametric Mann-Whitney test (A, B, C, E, L, M) or linear regression analysis (D, F-J). *** p < 0.001, **** p < 0.0001, ns, non-significant.

Article Snippet: ADAMTS13 antigen levels in the plasma of HC and TTP patients were measured using Human ADAMTS13 Quantikine ELISA Kit (R&D Systems) according to the manufacturer’s instructions with minor modifications: (1) diluted plasma samples of HC (1:50) and TTP patients (1:2) were used; (2) the range of the standard curve was broadened to 0.78125-100 ng/ml.

Techniques: Activity Assay, Concentration Assay, Derivative Assay, MANN-WHITNEY

Journal: Journal of cellular and molecular medicine

Article Title: The effect of ADAMTS13 on graft-versus-host disease.

doi: 10.1111/jcmm.18457

Figure Lengend Snippet: FIGURE 5 ADAMTS13 and VWF-A2 reduced donor-derived T cells in secondary lymphoid organs 24 h after bone marrow transplant. Mice were sacrificed 24 h post-transplant (n = 4). Cells were

Article Snippet: In some experiments, prior to adding Jurkat cells, histamine- stimulated HUVEC cells were treated with 1 μg/mL of recombinant human ADAMTS13 (6156- AD; R&D Systems) for 10 min at room temperature.

Techniques: Derivative Assay

Journal: Journal of cellular and molecular medicine

Article Title: The effect of ADAMTS13 on graft-versus-host disease.

doi: 10.1111/jcmm.18457

Figure Lengend Snippet: FIGURE 7 Role of VWF in the binding of T cells. (A) One hundred thousand Jurkat cells (immortalize malignant T lymphocytes) were incubated with HUVECs cells plated on 6-well plates. After 15 min incubation at 37°C, the number of adhered Jurkat cells in each well was counted using pictures taken with an inverted microscope and compared between histamine-stimulated and non-stimulated HUVECs. The effect of recombinant ADAMTS13 (1 mg/mL), recombinant VWF-A2 (1 mg/mL) and anti-αL antibodies (1:1000 dilution) on the number of adhered Jurkat cells to histamine- stimulated HUVECs were compared (n = 4, each experiment in triplicates). p-values were calculated using a one-way ANOVA test with Dunnett's multiple comparison correction compared to the number of adhered Jurkat cells to histamine-stimulated HUVECs. * p < 0.05. (B) One hundred thousand Jurkat cells were incubated in VWF-coated 96-wells plates for 15 min at 37°C. Jurkat cells, either pre-stimulated with 200 ng/mL of CCL21 or resting, were added to each well. The effect of recombinant ADAMTS13, recombinant VWF-A2 and anti-αL antibodies on the number of adhered CCL21-stimulated Jurkat cells to VWF was compared (n = 8, each experiment in triplicates). p-values were calculated using a one-way ANOVA test with Dunnett's multiple comparison correction compared to the number of adhered CCL21-stimulated Jurkat cells VWF. *p < 0.05.

Article Snippet: In some experiments, prior to adding Jurkat cells, histamine- stimulated HUVEC cells were treated with 1 μg/mL of recombinant human ADAMTS13 (6156- AD; R&D Systems) for 10 min at room temperature.

Techniques: Binding Assay, Incubation, Inverted Microscopy, Recombinant, Comparison

Journal: Journal of Clinical and Translational Hepatology

Article Title: ADAMTS13 Improves Hepatic Platelet Accumulation in Pyrrolizidine Alkaloids-induced Liver Injury

doi: 10.14218/JCTH.2024.00233

Figure Lengend Snippet: (A) Volcano plots of DEGs (PA-ILI group vs. control group). (B) Heatmap of the top 250 genes between PA-ILI mice (groups P1 & P2) and controls (groups C1 & C2). (C) Dot plots of seven GO biological process categories, including wound healing, hemostasis, coagulation, and platelet-related pathways. (D) Cnetplot displaying 57 DEGs assigned to the seven GO biological process categories. (E) Protein–protein interaction data of VWF from STRING, containing 11 nodes and 104 edges. (F) Venn diagram of 57 DEGs (red) and 11 VWF-related genes (blue) with three shared genes in the overlapping regions, including ADAMTS13. PA-ILI, pyrrolizidine alkaloids-induced liver injury; VWF, von Willebrand factor; DEG, differentially expressed genes; GO, Gene Ontology; STRING, Search Tool for the Retrieval of Interacting Genes/Proteins.

Article Snippet: Plasma ADAMTS13 levels were measured using a Human ADAMTS13 Quantikine ELISA kit (DADT130; R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions.

Techniques: Control, Coagulation

Journal: Journal of Clinical and Translational Hepatology

Article Title: ADAMTS13 Improves Hepatic Platelet Accumulation in Pyrrolizidine Alkaloids-induced Liver Injury

doi: 10.14218/JCTH.2024.00233

Figure Lengend Snippet: (A) Representative images (left) and quantification (right) of hepatic ADAMTS13 (red) in patients (n = 3). (B) Plasma level of ADAMTS13 in patients (leftmost). Peripheral platelet count and plasma ADAMTS13 concentration in mild and severe PA-ILI (middle). Relationship between peripheral platelet count and plasma ADAMTS13 in PA-ILI patients (rightmost). (C) Representative Western blots of ADAMTS13 in mouse plasma, lungs, and kidneys. (D) Hepatic mRNA level of ADAMTS13 in MCT-treated mice. (E) Representative Western blots of ADAMTS13 and apoptosis-related proteins. (F) Representative images (left) and quantification (right) of hepatic ADAMTS13 (red) in the mouse liver (n = 3). * p < 0.05, ** p < 0.01, **** p < 0.0001. PA-ILI, pyrrolizidine alkaloids-induced liver injury; DAPI, 4′,6-diamidino-2-phenylindole; MCT, monocrotaline.

Article Snippet: Plasma ADAMTS13 levels were measured using a Human ADAMTS13 Quantikine ELISA kit (DADT130; R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions.

Techniques: Clinical Proteomics, Concentration Assay, Western Blot

Journal: Journal of Clinical and Translational Hepatology

Article Title: ADAMTS13 Improves Hepatic Platelet Accumulation in Pyrrolizidine Alkaloids-induced Liver Injury

doi: 10.14218/JCTH.2024.00233

Figure Lengend Snippet: (A) Representative Western blots and quantification of plasma and hepatic ADAMTS13 protein. (B) Pathological changes in PA-ILI mice treated with rADAMTS13. (C) Measurement of hepatic necrosis (n = 5). (D) Representative images (left) and quantification (right) of hepatic platelets (CD41, red) and VWF deposition (green). * p < 0.05, ** p < 0.01, *** p < 0.001. PA-ILI, pyrrolizidine alkaloids-induced liver injury; VWF, von Willebrand factor; DAPI, 4′,6-diamidino-2-phenylindole; MCT, monocrotaline.

Article Snippet: Plasma ADAMTS13 levels were measured using a Human ADAMTS13 Quantikine ELISA kit (DADT130; R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions.

Techniques: Western Blot, Clinical Proteomics