Neutrophil-mediated oxidative stress and albumin structural damage predict COVID-19-associated mortality
- Research Department, Children’s Cancer Hospital, Egypt
- Infectious Disease Unit, Internal Medicine Department, Faculty of Medicine, Cairo University, Egypt
- Pharmacology Department, Faculty of Medicine, Assuit University, Egypt
- Pediatric Oncology Department, National Cancer Institute, Cairo University and Children's Cancer Hospital, Egypt
- Clinical pathology department, National Cancer Institute, Cairo University and Children's Cancer Hospital, Egypt
- Department of Intensive Care, Faculty of Medicine, Cairo University, Egypt
- Department of Pulmonary Medicine, Faculty of Medicine, Cairo University, Egypt
- Department of Chest Diseases, Faculty of Medicine, Cairo University, Egypt
- Division of Geriatric Medicine, Department of Medicine, Vanderbilt University Medical Center; and VATennessee Valley Geriatric Research, Education and Clinical Center (GRECC), United States
Figures
Figure 1
Probing structual changes of serum albumin through spin labeling EPR spectroscopy.
(A) HSA crystal structure containing seven copies of stearic acid. (B) Representative EPR spectra of free and HSA-bound 5-DSA (B) and 16-DSA (C) in whole blood from the same COVID-19 recovered …
Figure 2
Hematologic cellular counts and neutrophil-ROS levels reflect severity and mortality in COVID-19 patients.
(A) Representative flow cytometric diagrams comparing morphologic, hematologic, and ROS levels in control (representative of n=9; upper row), Sev-R (representative of n=10; middle row), and Sev-D …
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Figure 2—source data 1
Raw source data for Figure 2B-E.
- https://cdn.elifesciences.org/articles/69417/elife-69417-fig2-data1-v2.xlsx
Figure 3
Hydrogen peroxide levels in plasma and neutrophils reflect mortality in COVID-19 patients.
Catalase was used to specifically and quantitatively determine levels of hydrogen peroxide in identical plasma volumes collected from control (n=11), Sev-R (n=16), and Sev-D (n=23) groups. (A) …
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Figure 3—source data 1
Raw polarographic data for released oxygen (A), calibration curve (B), and calculated plasma hydrogen peroxide levels in all groups (C).
- https://cdn.elifesciences.org/articles/69417/elife-69417-fig3-data1-v2.xlsx
Figure 4
EPR spectroscopic analyses of HSA-fatty acid binding reveal strong dependence of binding on mortality in COVID-19 patients.
(A) Albumin level in plasma of control (n=8), Sev-R (n=16), and Sev-D (n = 23) groups showed that both survivors and non-survivors COVID-19 patients exhibit statistically significant …
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Figure 4—source data 1
Determined biochemical and biophysical EPR parameters used for statistical comparisons.
- https://cdn.elifesciences.org/articles/69417/elife-69417-fig4-data1-v2.xlsx
Figure 5
COVID-19-associated impairment of HSA transport function.
Transport function of HSA is assessed through the apparent kinetics of fatty acid uptake by following the rise in both strongly (A) and weakly (B) bound components immediately after mixing SLFA with …
Figure 6
Associations of oxidative stress measures and biophysical parameters with mortality outcome in COVID-19 subjects.
Linear correlations between plasma levels of hydrogen peroxide and EPR-calculated parameters pertaining to protein packing order parameter (A, B), fatty acid mobility τc (C), and S/W ratio (D) in …
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Figure 6—source data 1
Raw data for correlation analyses and for Kaplan-Meier analyses.
- https://cdn.elifesciences.org/articles/69417/elife-69417-fig6-data1-v2.xlsx
Tables
Table 1
Demographic and clinical characteristics of the studied subjects.
| Sev-R | Sev-D | Tukey 95% CI | p | |
|---|---|---|---|---|
| n | 16 | 23 | ||
| Age (mean ± SD) | 60.7±9.5 | 67.8±13.2 | 4.1–17.8 | 0.09 |
| Male | 56.25% | 63.16% | 0.677† | |
| sO2 (mean ± SD) | 82.1±18.7 | 76.1±18.6 | –20.5 to 8.4 | 0.40 |
| Hypertension | 12.5% | 42.1% | 0.053† | |
| Diabetes | 25% | 75% | 0.08† | |
| Cardiovascular disease | 0% | 15.8% | 0.10† | |
| Cancer | 0% | 10.5% | 0.18† | |
| Bronchial asthma | 6.25% | 10.5% | 0.65† | |
| ACE inhibitors | 0% | 7.14% | 0.47† | |
| ARBs | 9.09% | 7.14% | 0.85† | |
| calcium channel blocker | 14.28% | 7.14% | 0.60† | |
| Beta blockers | 0% | 7.14% | 0.47† | |
| Diuretics | 0% | 7.14% | 0.47† | |
| Sulphonylurea | 14.28% | 21.43% | 0.69† | |
| Other oral hypoglycemic | 0% | 21.43% | 0.18† | |
| Insulin | 42.85% | 21.43% | 0.30† | |
| Anticoagulant | 57.14% | 50.0% | 0.76† | |
| Steroids | 71.42% | 64.28% | 0.74† | |
| Hydroxychloro-quine | 14.28% | 7.14% | 0.60† | |
| IL-6 receptor antibody | 28.57% | 21.42% | 0.72† | |
| Proton-pump inhibitor | 28.57% | 42.85% | 0.52† | |
| Azithromycin | 14.28% | 28.57% | 0.47† | |
| Cephalosporin | 42.85% | 21.43% | 0.30† | |
| Carbapenem | 42.85% | 42.85% | 1.0† | |
| Oxazolidinone | 42.85% | 28.57% | 0.51† | |
| Fluoro-quinolone | 42.85% | 21.43% | 0.30† | |
| Nitrofuran | 14.28% | 0% | 0.15† | |
| Remdesivir | 14.28% | 28.57% | 0.47† | |
| Ivermectin | 0% | 28.57% | 0.11† |
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sO2, blood oxygen saturation level; ACE, angiotensin-converting enzyme; ARB, angiotensin II receptor blocker; IL-6, interleukin-6. † p values obtained through Pearson’s χ2 test.
Table 2
Laboratory parameters of the current study patients.
WBC, white blood cell; INR, international normalized ratio; CRP, high-sensitivity C-reactive protein; ICU,intensive care unit; PLT, platelet; ALT, alanine transaminase; AST, aspartate transaminase. …
| Sev-R (mean ± SD) | Sev-D (mean ± SD) | Tukey 95% CI | p | |
|---|---|---|---|---|
| WBCs (×103 /ml) | 10.6±4.0 | 13.9±8.0 | –1.47 to 8.06 | 0.17 |
| Platelets (×106 /ml) | 260±75.5 | 213.7±115.8 | –116.7 to 24.2 | 0.19 |
| INR | 1.29±0.56 | 1.23±0.23 | –0.38 to 0.26 | 0.69 |
| CRP (mg/L) | 51.19±54.4 | 103.77±86.3 | –2.35 to 107.5 | 0.06 |
| D-dimer (mg/ml) | 1.47±1.9 | 3.17±3.56 | –0.55 to 3.96 | 0.13 |
| IL-6 (pg/ml) | 314.1±527 | 325.3±619 | –591 to 614 | 0.97 |
| Ferritin | 922.6±565 | 1078±578 | –281 to 594 | 0.47 |
| Albumin (g/ml) | 31.47±7.95 | 26.97±5.1 | –8.7 to –0.26 | 0.038 |
| Hemoglobin (g/dl) | 12.26±2.0 | 12.16±2.0 | –1.53 to –1.33 | 0.97 |
| ALT (U/L) | 33.64±24.15 | 46.5±37.2 | –10.37 to 36.13 |
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ALT, alanine transaminase; AST, aspartate transaminase; CRP, high-sensitivity C-reactive protein; ICU,intensive care unit; PLT, platelet; INR, international normalized ratio; WBC, white blood cell. The Tukey’scalculated p-values as well as upper and lower 95% confidence levels for the Sev-R vs. Sev-D means’comparisons are given.
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | CD-42b-PE- (Mouse monoclonal) | Beckman Coulter Life Sciences | Cat#IM1417URRID: AB_2893282 | FACS (1 µl per test) |
| Antibody | CD14-PC7- (Mouse monoclonal) | Beckman Coulter Life Sciences | Cat#A22331RRID: AB_10639528 | FACS (1 µl per test) |
| Antibody | CD66b-APC-Alexa Fluor 750- (Mouse monoclonal) | Beckman Coulter Life Sciences | Cat#B08756RRID:AB_2893284 | FACS (1 µl per test) |
| Antibody | CD3-ECD- (Mouse monoclonal) | Beckman Coulter Life Sciences | Cat#IM2705URRID: MGI:3850637 | FACS (1 µl per test) |
| Other | 2′,7′-dichlorofluorescein-diacetate | Sigma-Aldrich | Cat#D6883 | Imaging (30 μM)FACS (20 μM) |
| Other | DAPI | Thermo Fisher Scientific | Cat#62249 | Imaging (6.15 µg/ml) |
| Other | 2-(3-carboxypropyl)-4,4-dimethyl-2-tridecyl-3-oxazolidinyloxy (5–130 doxyl-stearic acid, 5-DSA) | Sigma-Aldrich | Cat#253618 | EPR (0.26 mm) |
| Other | 2-(14-carboxytetradecyl)-2-ethyl-4,4-dimethyl-3-oxazolidinyloxy (16–131 doxyl-stearic acid, 16-DSA) | Sigma-Aldrich | Cat#253596 | EPR (0.26 mm)EPR/ethanol experiments (0.52 mm) |
Additional files
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Supplementary file 1
Calculated biophysical EPR spectral parameters for all groups.
- https://cdn.elifesciences.org/articles/69417/elife-69417-supp1-v2.pptx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/69417/elife-69417-transrepform1-v2.docx
