Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: Inhibitors. The manufacturers and working concentrations of the inhibitors used in this study are listed.

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Concentration Assay, Control

Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: CD34 + cells were cultured with the inhibitors at the concentrations indicated in the graphs for 48 and 72 hours (Mean cell counts with standard deviation).

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Cell Culture, Standard Deviation

Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: CD34 + cells derived from healthy volunteers were differentiated for 14 days and subsequently lysed. The lysates were incubated with or without inhibitors, following the manufacturer's protocol. BAY-677, the inactive analog of inhibitor BAY-678, was included as a negative control. Neutrophil elastase proteolytic activity was measured using the EnzChek elastase assay kit and quantified with a fluorescence microplate reader (Mean Fluorescence Units with standard deviation).

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Derivative Assay, Incubation, Negative Control, Activity Assay, Fluorescence, Standard Deviation

Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: Healthy volunteer and patient derived CD34+ cells were differentiated for 14 days in the presence or absence of inhibitors. The resultant cells were labeled with antibodies to CD14, CD66b, CD11b, and CD15 surface markers and analyzed using flow cytometry. A. Representative experiment histograms are shown. The proportion of CD14 + /CD66b + and CD11b + /CD15 + positive cells in quadrant 2 are indicated. B. Graphical representation of the percentage of CD66b + /CD14 + and CD11b + /CD15 + cellular subsets of the patients' cells after the addition of NE inhibitors. For each individual experiment, the percentage of cells with a mature phenotype after addition of inhibitor was divided by the percentage measured when only the vehicle control was added and this ratio was plotted. Data from 5 different patients, represented in at least two different experiments. Each individual patient has a different symbol. C. Cell cytospins stained with Kwik-Diff (eosin/methylene blue) were imaged using a Nikon digital camera. Cell differentiation was evaluated at 400x magnification by light microscope. Representative experiments showing the effect of MK0339 and brensocatib inhibitors on healthy volunteer and patient cells are shown.

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Derivative Assay, Labeling, Flow Cytometry, Control, Staining, Cell Differentiation, Light Microscopy

Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: Molecular docking simulations for NE inhibitors were conducted using the MOE software package. The simulations utilized ELANE G214R, P139L, and wild-type models of neutrophil elastase. The images captured from the simulations display the binding sites of the inhibitors MK0339, sivelestat, GW311616, and BAY-678 with the wild-type NE. The inhibitor molecules are highlighted in color and indicated by arrows.

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Software, Binding Assay

Journal: Journal of cellular immunology

Article Title: Evaluation of Neutrophil Elastase Inhibitors as Potential Therapies for ELANE Associated Neutropenia

doi: 10.33696/immunology.6.208

Figure Lengend Snippet: Molecular docking simulation analysis. Molecular docking simulations for NE inhibitors were conducted using the MOE software package. The simulations utilized ELANE G214R, P139L, and wild-type models of neutrophil elastase. The stability of each inhibitor-protein complex was assessed using the free energy of binding (S score in kcal/mol), providing an approximation of binding stability.

Article Snippet: NE inhibitors: Sivelestat (HY-17443), GW311616 (HY-15891), BAY-678 (HY-111457A) with its inactive control, BAY-677 (HY-111457) and DPP1 inhibitor Brensocatib (HY-101056) were purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Software, Binding Assay