adamts13 antigen  (Elabscience Biotechnology)

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: ADAMTS13 reduced GVHD in mice. ADAMTS13 treatment (at 300 μg/kg) reduced mortality and morbidity in acute GVHD in mice. (A) Survival was monitored daily for 4 weeks, with results from three independent experiments ( p = 0.0025, n = 15 for each group). (B) Mice were sacrificed on Day 7 post‐transplant (five mice per group). The top and lower panels represent the skin, small intestine and liver of mice treated with PBS control and ADAMTS13, respectively. Arrows and asterisks indicate the histopathologic changes in the skin (inflammatory and apoptotic cells in hair follicles), intestine (necrosis in the crypts), liver (lymphoid infiltration in portal tracts and epithelial damage of bile duct) and lung (lesions with inflammatory infiltration). (C) Average GVHD score of skin, intestine, liver and lung. Results are shown as mean ± SD ( t ‐test; n = 5 in each group). * p < 0.05.

Article Snippet: Recombinant ADAMTS13 was purchased from Bio‐techne/Tocris (Minneapolis MN), and recombinant VWF‐A2 protein was produced, as described previously.

Techniques: Control

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: ADAMTS13 altered the number and polarization of donor‐derived T cells in mice. Mice were sacrificed on Day 7 post‐transplant (three mice per group). Cells were collected from the spleen (SP) and peripheral lymph nodes (LN) and then stained with CD3, CD4, CD8, H‐2D b , Foxp3 and CD25 antibodies. (A) The number of donor‐derived T‐cell subsets recovered from each tissue, CD4 + /H‐2D b+ , CD8 + /H‐2D b+ and CD4 + /CD25 + /Foxp3 + /H‐2D b+ (Treg) were determined by FACS. (B) Intracellular expression of IL‐2, IL‐4, IL17, IFNƔ and TNF‐α in donor‐derived T cells was compared between ADAMTS13‐ and PBS‐treated recipient mice. Results were calculated as the mean and SD of at least three independent experiments. p ‐values were calculated using a t ‐test comparing ADAMTS13‐treated and PBS control mice. Results are shown as bar graphs representing means ± SD. * p < 0.05.

Article Snippet: Recombinant ADAMTS13 was purchased from Bio‐techne/Tocris (Minneapolis MN), and recombinant VWF‐A2 protein was produced, as described previously.

Techniques: Derivative Assay, Staining, Expressing, Control

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: 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 collected from the spleen (SP), peripheral lymph nodes (PLN) and mesenteric lymph nodes (MLNs) and then stained with H‐2D b , CD3, CD4, CD8, CD25 and Foxp3 antibodies. The number of donor‐derived T cells and their subsets recovered from each tissue are shown. (A) The total number of donor‐derived T cells (CD3 + H‐2D b+ ), (B) CD4+ (CD4 + / CD3 + /H‐2D b+ ), (C) CD8+ (CD8 + /CD3 + /H‐2D b+ ) and (D) Tregs (CD4 + /CD3 + /CD25 + /Foxp3 + /H‐2D b+ ) was determined by FACS. Results are shown as bar graphs representing mean ± SD. p‐ values were calculated using a t ‐test comparing ADAMTS13‐treated or VWF‐A2‐treated and PBS‐treated (control) mice. * p < 0.05.

Article Snippet: Recombinant ADAMTS13 was purchased from Bio‐techne/Tocris (Minneapolis MN), and recombinant VWF‐A2 protein was produced, as described previously.

Techniques: Derivative Assay, Staining, Control

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: ADAMTS13 and VWF‐A2 did not impact T‐cell proliferation in vitro. CFSE‐labelled C57BL/6 responder cells were cocultured at a ratio of 1:1 with irradiated Balb/C stimulator cells for 7 days at 37°C. Primary T‐cell proliferation percentage was assessed by CFSE dye dilution following sequential gating on antibody‐stained (A) CD4 + or (B) CD8 + T cells. The number of alive (C) CD4 + and (D) CD8 + T cells was averaged from three independent experiments in MLR. Results are shown as bar graphs representing mean ± SD. p‐ values were calculated using a t ‐test. * p < 0.05. NC: negative control, C57BL/B6 responder splenocytes only (unstimulated T cells). PC: positive control, C57BL/B6 responder splenocytes cocultured with irradiated Balb/C stimulator splenocytes at 1:1 ratio.

Article Snippet: Recombinant ADAMTS13 was purchased from Bio‐techne/Tocris (Minneapolis MN), and recombinant VWF‐A2 protein was produced, as described previously.

Techniques: In Vitro, Irradiation, Staining, Negative Control, Positive Control

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: 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‐α L 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: Recombinant ADAMTS13 was purchased from Bio‐techne/Tocris (Minneapolis MN), and recombinant VWF‐A2 protein was produced, as described previously.

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

Journal: The New England journal of medicine

Article Title: Recombinant ADAMTS13 for Immune Thrombotic Thrombocytopenic Purpura

doi: 10.1056/NEJMoa2402567

Figure Lengend Snippet: Panel A shows serial half-life modeling for rADAMTS13 over 14 non-consecutive days using fitted one-phase exponential decay curves based on ADAMTS13 activity levels measured at 0, 1, 4, 8, and 12 hours after administration (left) as well as the computed ADAMTS13 activity half-life by treatment day (right). The red line and data point represent the first dose, while the blue line and data point represent the 41st dose. The dotted line in the left panel represents 10% ADAMTS13 activity level, the diagnostic cutoff for severe deficiency. The results of simple linear regression analysis are shown in the right panel. Panel B depicts serial ADAMTS13 activity measurements between days 0 and 30. The vertical red arrow and mark denote initiation of intravenous rADAMTS13 treatment. Similarly, ADAMTS13 autoantibody was measured serially via two independent methods (Panel C), including titer in Bethesda-like units (BU) evaluated by FRETS-VWF73 assay (blue) and plasma anti-ADAMTS13 antibody concentration, measured by sandwich ELISA (black). The gray bar denotes periods during which rADAMTS13 (80 IU/kg) was dosed every 12 hours, while the black bar represents dosing every 24 hours. Immunoblotting of patient plasma for ADAMTS13 at 3 timepoints relative to the first dose of rADAMTS13 is shown Panel D alongside normal pooled plasma (NPP) and purified rADAMTS13 in buffer. Corresponding levels of ADAMTS13 antigen (assessed by densitometry) and activity (assessed by FRETS-VWF73 assay) are noted below the image. Assessments of total ADAMTS13 antigen and autoantibody-bound plasma ADAMTS13 levels are shown in Panel E. Sandwich ELISA assays were performed on patient plasma at the indicated times after rADAMTS13 administration to detect total plasma ADAMTS13 (red) and autoantibody-bound ADAMTS13 in the form of circulating immune complexes (CIC) (black). ADAMTS13 CIC concentration is expressed in arbitrary units (AU), while total plasma ADAMTS13 is expressed in ng/ml. Time 0 samples were drawn immediately after daily plasma exchange treatment and prior to administration of rADAMTS13. Data represents the mean (± SD) of four consecutive dosing cycles. The horizontal red dotted line represents the mean baseline plasma ADAMTS13 antigen level at time 0 (225.9 ng/ml).

Article Snippet: Plasma ADAMTS13 antigen was quantified by sandwich ELISA (ProteinTech product #KE00286).

Techniques: Activity Assay, Diagnostic Assay, Clinical Proteomics, Concentration Assay, Sandwich ELISA, Western Blot, Purification