Journal: Pathogens and Immunity

Article Title: Immune Dysregulation in Acute SARS-CoV-2 Infection

doi: 10.20411/pai.v7i2.537

Figure Lengend Snippet: Analysis of ACE2 inhibition by anti-S1(A) and anti-RBD (B) IgG antibodies by disease severity. Quantitative plasma anti-S had a correlation of r = 0.885 ( P < 0.001) and anti-RBD had a correlation of r = 0.75 ( P < 0.001) to ACE2 neutralization.

Article Snippet: The detection antibody used for this assay is R&D Systems Human ACE2 Biotinylated Antibody, (catalog BAF933).

Techniques: Inhibition, Clinical Proteomics, Neutralization

Journal: Pathogens and Immunity

Article Title: Immune Dysregulation in Acute SARS-CoV-2 Infection

doi: 10.20411/pai.v7i2.537

Figure Lengend Snippet: Proinflammatory dysregulation between cytokine microenvironment and antibody neutralization capacity. Correlation analysis of inflammatory marker plasma concentration and anti-spike antibody ACE2 neutralization disease severity. Elevations in ICAM ( P < 0.0001), IL-1β ( P = 0.0233), IL-4 ( P < 0.0001), IL-6 ( P = 0.0001), TNFα ( P = 0.001), and Syndecan ( P = 0.00) all showed statistically significant correlations to antibody neutralization capacity amongst PCR-positive COVID-19 disease cohorts.

Article Snippet: The detection antibody used for this assay is R&D Systems Human ACE2 Biotinylated Antibody, (catalog BAF933).

Techniques: Neutralization, Marker, Clinical Proteomics, Concentration Assay

Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Serum ACE2 activity is significantly correlated with SBP in stroke-alert patients and healthy young adults, but not AIS patients. Correlation graphs of ACE2 activity and SBP among stroke-alert patients (a) and healthy young adults (b) as compared to stroke patients (c). Young adult blood plasma samples in panel (b) were from a biorepository established by Wegman et al., which were obtained from research participants undergoing baseline measurements. (d) Correlation graph of ACE activity and mRS at discharge from hospital among AIS patients. ACE2: angiotensin converting enzyme 2; AIS: acute ischemic stroke; mRS: modified Rankin score; RFU: relative fluorescence unit; SBP: systolic blood pressure.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay, Clinical Proteomics, Modification, Fluorescence

Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Activity of ACE2 and ACE in serum is altered following stroke. For human serum, bar graphs are means ± SEM and represent enzyme activity levels of ACE2 (a) and ACE (c) from control, stroke-alert, or AIS patients at an average of 3.6 hours and again at 3 days after stroke. Individual differences and means ± SEM in ACE2 (b) and ACE (d) are shown. * P <0.05 versus control and † P <0.05 versus stroke-alert. ‡ P <0.05 versus AIS <6 hours. ACE: angiotensin converting enzyme; ACE2: angiotensin converting enzyme 2; AIS: acute ischemic stroke; RFU: relative fluorescence unit.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay, Control, Fluorescence

Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Predictors of acute ischemic stroke by multiple linear regression analysis.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant

doi: 10.3390/ijms241814140

Figure Lengend Snippet: ACE2, HS, and SDC expression profiles of 293T and 293T-ACE2 cells. ACE2, HS, and SDC expression in 293T and 293T-ACE2 cells were assessed using fluorescently labeled specific antibodies with imaging flow cytometry. ( A , B ) ACE2 and HS expression profile of 293T and 293T-ACE2 cells. The representative flow cytometry histograms and cellular images show the ACE2 and HS expression of 293T and 293T-ACE2 cells treated with the fluorescent ACE2 and HS antibodies. Scale bar = 20 μm. ( C ) SDC expression profile of 293T and 293T-ACE2 cells. The representative flow cytometry histograms and fluorescent cellular images show the SDC expression of 293T cells and 293T-ACE2 cells treated with the APC-labeled respective SDC antibodies. Scale bar = 20 μm. ( D – F ) Detected ACE2, HS, and SDC expression values were normalized to those of 293T cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Article Snippet: ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol.

Techniques: Expressing, Labeling, Imaging, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant

doi: 10.3390/ijms241814140

Figure Lengend Snippet: Cellular uptake of WT SCV2, Delta, and Omicron variants into 293T- and ACE2-overexpressing 293T-ACE2 cells. The cells were exposed to 1 MOI of heat-inactivated WT SCV2, Delta, and Omicron variants for 4 h at 37 °C. After incubation, the cells were washed, trypsinized, fixed, permeabilized, and treated with a primary SARS-CoV-2 spike (1000–1200 aa), followed by a fluorescently labeled (AF 633) secondary antibody. Cellular uptake was then analyzed with imaging flow cytometry. ( A – D ) Representative flow cytometry histograms and brightfield (BF) and fluorescent cellular images showing the intracellular fluorescence of the virus-exposed 293T and 293T-ACE2 cells. Scale bar = 20 μm. ( E ) Detected fluorescence intensities were normalized to WT SCV2-treated 293T cells as standards. The bars represent the mean + SEM of five independent experiments. Experimental data are presented as dots. Statistical significance was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.

Article Snippet: ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol.

Techniques: Incubation, Labeling, Imaging, Flow Cytometry, Fluorescence, Virus