Background
Methods
Patients and controls
Blood samples
Cytokines and P-selectin
Cell-free DNA
Statistical analysis
Results
Patients and controls
Viral phase (1–9 days) N = 87 | Early inflammatory (10–16 days) N = 53 | Late inflammatory (> 16 days) N = 40 | |
---|---|---|---|
Days from symptom onset | 6.00 [5.00; 8.00] | 12.00 [11.00; 14.00] | 20.00 [18.00; 24.25] |
A | ||||
---|---|---|---|---|
CCDC scale | Mild-moderate | Severe | Critical | Total |
Patients (n) | 51 | 15 | 27 | 93 |
WHO OS | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | Total |
---|---|---|---|---|---|---|---|---|---|
Patients (n) | 0 | 0 | 7 | 41 | 20 | 1 | 5 | 19 | 93 |
B | ||||
---|---|---|---|---|
Illness onset | 1–9 days | 10–16 days | ≥ 17 days | Total |
93 patient samples (n) | 97 | 59 | 45 | 201 |
72 patient samples (n) | 87 | 53 | 40 | 180 |
Patients N = 93 | Controls N = 55 | p | ||
---|---|---|---|---|
Sex | Female | 51 (54.84%) | 30 (54.55%) | 1.000 |
Male | 42 (45.27%) | 25 (45.45%) | ||
Age (years) | 69.00 [55.00; 78.00] | 71.00 [57.00; 79.00] | 0.441 | |
Hypertension | 45 (48.39%) | 17 (30.91%) | 0.056 | |
Diabetes | 28 (30.11%) | 12 (21.82%) | 0.365 | |
Obesity | 13 (13.98%) | 4 (7.27%) | 0.332 | |
Heart disease | 14 (15.05%) | 6 (10.91%) | 0.643 | |
Chronic respiratory disease | 16 (17.20%) | 3 (5.45%) | 0.070 | |
Liver disease | 3 (3.23%) | 0 (0.00%) | 0.295 | |
Renal disease | 8 (8.60%) | 6 (10.91%) | 0.863 | |
Dementia | 18 (19.35%) | 4 (7.27%) | 0.079 | |
Dyslipidemia | 17 (18.28%) | 17 (30.91%) | 0.118 |
All samples | Patients N = 201 | Controls N = 55 | p |
---|---|---|---|
TNF-α (pg/mL) | 40.02 [34.83; 47.41] | 41.07 [38.56; 44.99] | 0.156 |
IL-8 (pg/mL) | 54.29 [40.43; 89.22] | 56.21 [43.26; 107.75] | 0.412 |
IL-1β (pg/mL) | 55.90 [47.80; 66.92] | 59.91 [54.04; 62.24] | 0.238 |
IFN-γ (pg/mL) | 145.84 [123.72; 176.59] | 134.60 [128.65; 142.43] | 0.004 |
IL-17A (pg/mL) | 19.40 [14.73; 23.90] | 21.31 [18.48; 21.31] | 0.204 |
P-Selectin (ng/mL) | 55.48 [42.12; 73.30] | 77.44 [67.55; 84.99] | < 0.001 |
G-CSF (pg/mL) | 143.97 [123.70; 159.45] | 130.47 [124.32; 138.97] | < 0.001 |
IL-6 (pg/mL) | 20.41 [4.34; 51.62] | 7.80 [5.15; 10.42] | 0.004 |
cfDNA (ng/mL) | 7.87 [4.45; 14.40] | 2.56 [1.71; 3.61] | < 0.001 |
1st samples | Patients N = 93 | Controls N = 55 | p |
---|---|---|---|
TNF-α (pg/mL) | 38.51 [32.58; 45.34] | 41.07 [38.56; 44.99] | 0.013 |
IL-8 (pg/mL) | 47.45 [33.71; 66.16] | 56.21 [43.26; 107.75] | 0.010 |
IL-1β (pg/mL) | 51.66 [43.09; 59.90] | 59.91 [54.04; 62.24] | < 0.001 |
IFN-γ (pg/mL) | 134.40 [119.97; 159.07] | 134.60 [128.65; 142.43] | 0.846 |
IL-17A (pg/mL) | 18.48 [10.79; 22.80] | 21.31 [18.48; 21.31] | 0.001 |
P-Selectin (ng/mL) | 52.21 [35.63; 71.42] | 77.44 [67.55; 84.99] | < 0.001 |
G-CSF (pg/mL) | 142.57 [120.38; 162.04] | 130.47 [124.32; 138.97] | 0.041 |
IL-6 (pg/mL) | 30.52 [6.26; 62.51] | 7.80 [5.15; 10.42] | < 0.001 |
cfDNA (ng/mL) | 7.68 [4.23; 14.49] | 2.56 [1.71; 3.61] | < 0.001 |
Cytokines and tissue damage markers
Severe and Critical | Moderate | p | ||
---|---|---|---|---|
CCDC scale | IL-6 (pg/mL) | 32.44 [5.84; 104.03] | 9.00 [2.76; 33.84] | < 0.001 |
cfDNA (ng/mL) | 9.44 [6.15; 20.19] | 6.21 [3.46; 10.44] | < 0.001 | |
WHO OS | IL-6 (pg/mL) | 32.44 [5.69; 99.94] | 9.00 [2.90; 33.14] | < 0.001 |
cfDNA (ng/mL) | 9.08 [5.54; 19.38] | 6.88 [3.61; 10.73] | 0.001 |
Critical | Moderate and Severe | p | ||
---|---|---|---|---|
CCDC scale | IL-6 (pg/mL) | 59.48 [21.37; 175.20] | 10.06 [3.10; 34.21] | < 0.001 |
cfDNA (ng/mL) | 14.11 [7.13; 22.67] | 7.01 [4.01; 10.93] | < 0.001 |
Survivors | Non-survivors | p | ||
---|---|---|---|---|
Total samples (n = 201) | IL-6 (pg/mL) | 11.14 [3.49; 39.23] n = 158 | 52.49 [21.37; 152.90] n = 43 | < 0.001 |
cfDNA (ng/mL) | 7.06 [4.12; 11.40] n = 158 | 14.98 [7.42; 25.74] n = 43 | < 0.001 | |
1st sample (n = 93) | IL-6 (pg/mL) | 25.91 [5.77; 51.55] n = 74 | 66.91 [38.38; 162.70] n = 19 | 0.001 |
cfDNA (ng/mL) | 6.87 [4.01; 10.81] n = 74 | 14.98 [7.73; 22.51] n = 19 | 0.002 |
Viral phase (1) N = 87 | Early inflammatory (2) N = 53 | Late inflammatory (3) N = 40 | p | p 1 vs 2 | p 1 vs 3 | p 2 vs 3 | |
---|---|---|---|---|---|---|---|
Leukocytes/mm3 | 7000 [5300;9050] | 8000 [6400;11400] | 8200 [6450;11800] | 0.034 | 0.092 | 0.056 | 0.582 |
Neutrophils/mm3 | 5100 [3350;6950] | 6000 [4400;8700] | 6050 [4100;8225] | 0.069 | 0.119 | 0.119 | 0.874 |
Lymphocytes/mm3 | 1000 [650;1400] | 1000 [700;1600] | 1150 [675;1900] | 0.705 | 0.858 | 0.858 | 0.858 |
N/L ratio | 5.12 [2.71;9.11] | 6.02 [2.61;14.35] | 4.96 [2.74;11.03] | 0.700 | 0.836 | 0.836 | 0.836 |
Platelet/mm3 | 234,000 [167500;285500] | 278,000 [197000;367000] | 227,000 [186000;278250] | 0.030 | 0.028 | 0.667 | 0.114 |
CRP (mg/L) | 50.00 [17.70;108.05] | 25.10 [9.40;74.60] | 17.00 [3.08;51.45] | 0.004 | 0.032 | 0.011 | 0.261 |
PCT (ng/mL) | 0.08 [0.05;0.16] | 0.07 [0.04;0.12] | 0.08 [0.04;0.23] | 0.415 | 0.564 | 0.620 | 0.620 |
LDH (U/L) | 294.50 [223.25;385.25] | 274.00 [201.00;345.00] | 232.00 [173.00;327.25] | 0.127 | 0.362 | 0.135 | 0.362 |
D-Dimer (µg/L) | 865.00 [434.00;1464.00] | 855.50 [495.25;1597.75] | 698.50 [345.50;1951.25] | 0.776 | 0.998 | 0.848 | 0.848 |
Ferritin (ng/mL) | 445.00 [219.80;620.90] | 647.50 [321.67;945.40] | 397.00 [248.50;726.10] | 0.107 | 0.103 | 0.808 | 0.343 |
TNF-α (pg/mL) | 36.40 [32.80;43.59] | 44.28 [38.81;49.81] | 41.11 [37.36;48.52] | < 0.001 | 0.001 | 0.010 | 0.381 |
IL-8 (pg/mL) | 52.01 [34.96;68.72] | 55.52 [45.64;91.01] | 77.32 [50.36;106.06] | 0.002 | 0.048 | 0.002 | 0.108 |
IL-1β (pg/mL) | 50.63 [43.09;61.88] | 60.12 [54.10;69.95] | 64.34 [53.61;69.23] | < 0.001 | < 0.001 | < 0.001 | 0.954 |
IFN-ʏ (pg/mL) | 131.17 [116.20;158.99] | 164.89 [143.24;179.26] | 157.76 [134.66;188.20] | < 0.001 | < 0.001 | 0.001 | 0.578 |
IL-17A (pg/mL) | 18.48 [10.79;22.80] | 21.69 [18.71;28.26] | 23.90 [15.45;24.77] | < 0.001 | < 0.001 | 0.001 | 0.714 |
P-Selectin (ng/mL) | 47.42 [34.91;60.86] | 65.42 [49.03;80.14] | 64.95 [51.05;79.88] | < 0.001 | < 0.001 | < 0.001 | 0.842 |
G-CSF (pg/mL) | 139.08 [120.98;160.64] | 147.85 [129.84;162.19] | 148.79 [132.43;157.64] | 0.478 | 0.488 | 0.488 | 0.964 |
IL-6 (pg/mL) | 29.11 [6.49;58.41] | 10.38 [2.59;31.52] | 10.06 [3.20;50.89] | 0.024 | 0.021 | 0.190 | 0.552 |
cfDNA (ng/mL) | 6.40 [3.61;13.99] | 7.87 [5.34;14.65] | 10.53 [5.56;17.30] | 0.041 | 0.200 | 0.043 | 0.200 |
SaO2/FiO2 | 444.50 [365.00;460.75] | 429.00 [339.00;457.00] | 330.50 [170.75;467.75] | 0.832 | 0.938 | 0.938 | 0.938 |
SaO2 | 94.00 [88.50;96.00] | 90.00 [87.00;95.00] | 93.00 [88.00;95.00] | 0.035 | 0.057 | 0.109 | 0.466 |
Survivors N = 74 | Non-survivors N = 19 | p | |
---|---|---|---|
Leukocytes/mm3 | 7200 [5425;8775] | 9800 [6600;13150] | 0.028 |
Neutrophils/mm3 | 5050 [3725;6550] | 8400 [4950;11750] | 0.009 |
Lymphocytes/mm3 | 1200 [800;1600] | 700 [532.50;800] | 0.001 |
N/L ratio | 4.46 [2.51;7.72] | 13.50 [6.75;20.95] | 0.001 |
Platelet/mm3 | 245,000 [168750;279750] | 228,000 [192000;327000] | 0.675 |
CRP (mg/L) | 33.90 [11.70;95.10] | 104.10 [59.20;197.00] | 0.008 |
PCT (ng/mL) | 0.07 [0.04;0.11] | 0.14 [0.08;0.60] | 0.025 |
LDH (U/L) | 255.00 [208.50;366.50] | 323.00 [237.50;403.50] | 0.223 |
D-Dimer (µg/L) | 828.50 [471.75;1378.50] | 1533.50 [1009.75;2671.00] | 0.009 |
Ferritin (ng/mL) | 427.30 [176.00;623.90] | 491.70 [392.35;963.60] | 0.110 |
TNF-α (pg/mL) | 37.84 [31.76;44.02] | 40.91 [32.95;49.72] | 0.194 |
IL-8 (pg/mL) | 42.11 [33.18;56.48] | 61.83 [52.88;89.06] | 0.016 |
IL-1β (pg/mL) | 50.63 [43.09;58.71] | 53.75 [45.44;65.57] | 0.314 |
IFN-ʏ (pg/mL) | 132.22 [119.97;154.89] | 144.93 [117.45;166.73] | 0.737 |
IL-17A (pg/mL) | 15.67 [10.79;22.80] | 19.40 [13.35;22.53] | 0.287 |
P-Selectin (ng/mL) | 49.25 [35.63;71.35] | 53.59 [36.85;69.489] | 0.900 |
G-CSF (pg/mL) | 134.22 [118.12;153.99] | 160.09 [139.92;184.10] | 0.012 |
IL-6 (pg/mL) | 25.91 [5.77;51.55] | 66.91 [38.36;162.70] | 0.001 |
cfDNA (ng/mL) | 6.87 [4.01;10.81] | 14.98 [7.73;22.51] | 0.002 |
SaO2/FiO2 | 452.00 [425.50;462.00] | 380.00 [323.00;416.50] | 0.003 |
SaO2 (%) | 95.00 [90.00;97.00] | 85.00 [75.50;86.50] | < 0.001 |
Severity | cfDNA (ng/mL) | p |
---|---|---|
CCDC scale | < 0.001 | |
Moderate (N = 105) | 6.42 (3.49–10.48) | |
Severe (N = 38) | 7.47 (5.32–13.37) | |
Critical (N = 58) | 14.92 (7.18–24.68) | |
Critical vs. moderate | < 0.001 | |
Critical vs. severe | 0.004 | |
WHO OS | 0.001 | |
Moderate; N = 102 | 6.97 (3.76–10.83) | |
Severe/Critical; N = 99 | 9.36 (5.56–19.60) |
Mortality | cfDNA (ng/mL) | p |
---|---|---|
Non-survivors N = 39 | 0.021 | |
Viral (1–9 days); N = 17 | 7.87 [5.52–22.38] | |
Early inflammatory (10–16 days); N = 11 | 12.59 [7.88–26.36] | |
Late inflammatory (> 17 days); N = 11 | 30.34 [18.47–39.39] | |
Viral phase vs late inflammatory | 0.031 | |
Survivors N = 141 | 0.328 | |
Viral (1–9 days); N = 70 | 5.98 (3.12–11.45) | |
Early inflammatory (10–16 days); N = 42 | 7.11 (4.59–10.60) | |
Late inflammatory (> 17 days); N = 29 | 7.83 (4.65–12.18) |
Correlation cfDNA with IL-6 and LDH | r | p |
---|---|---|
IL-6 | ||
Viral (1–9 days) | 0.12, (− 0.104–0.332) | 0.294 |
Early inflammatory (10–16 days) | 0.468 (0.192–0.672) | 0.002 |
Late inflammatory (> 17 days) | − 0.005 (− 0.348–0.339) | 0.977 |
LDH | ||
Viral (1–9 days) | 0.296 (0.079–0.486) | 0.009 |
Early inflammatory (10–16 days) | 0.376 (0.115–0.589) | 0.006 |
Late inflammatory (> 17 days) | 0.294 (− 0.024–0.558) | 0.069 |
Viral phase (1–9 days) N = 97 | Survivors N = 78 | Non-survivors N = 19 | p |
---|---|---|---|
Neutrophils/mm3 | 4500 [3150;6600] | 7000 [4600;10550] | 0.011 |
Lymphocytes/mm3 | 1200 [800;1600] | 600 [450;800] | < 0.001 |
N/L ratio | 4.25 [2.10;7.49] | 13.50 [6.75;25.85] | < 0.001 |
CRP (mg/L) | 38.0 [12.33;93.83] | 95.00 [59.20;153.40] | 0.047 |
Ferritin (ng/mL) | 406.00 [176.00;569.50] | 558.90 [460.75;1046.17] | 0.015 |
IL-6 (pg/mL) | 27.27 [5.78;51.08] | 57.27 [31.73;98.75] | 0.039 |
cfDNA (ng/mL) | 6.11 [3.48;11.45] | 7.87 [5.08;22.21] | 0.037 |
Early inflammatory (10–16 days) N = 59 | Survivors N = 48 | Non-survivors N = 11 | p |
---|---|---|---|
Leukocytes/mm3 | 7500 [6475;8825] | 12,600 [8900;16250] | 0.004 |
Neutrophils/mm3 | 5600 [4275;7000] | 11,400 [7650;14550] | < 0.001 |
Lymphocytes/mm3 | 1200 [775;1800] | 700 [400;800] | 0.002 |
N/L ratio | 4.40 [2.39;9.06] | 17.14 [15.59;24.93] | < 0.001 |
CRP (mg/L) | 20.30 [5.65;41.20] | 72.25 [38.75;128.12] | 0.004 |
PCT (ng/mL) | 0.05 [0.04;0.11] | 0.12 [0.07;0.21] | 0.023 |
D-Dimer (µg/L) | 705.50 [469.50;1020.75] | 3256.50 [1292.25;4987.75] | 0.014 |
IL-6 (pg/mL) | 5.28 [1.50;23.10] | 49.12 [26.78;214.49] | 0.001 |
cfDNA (ng/mL) | 7.11 [4.46;9.75] | 12.59 [7.88;26.36] | 0.008 |
Late inflammatory (> 16 days) N = 45 | Survivors N = 32 | Non-survivors N = 13 | p |
---|---|---|---|
Leukocytes/mm3 | 7400 [6300;10250] | 9800 [8700;18700] | 0.003 |
Neutrophils/mm3 | 5250 [4075;6625] | 8400 [6500;16400] | 0.001 |
Lymphocytes/mm3 | 1300 [800;2300] | 600 [400;1100] | 0.002 |
N/L ratio | 3.82 [2.55;6.19] | 15.50 [7.64;41.00] | < 0.001 |
CRP (mg/L) | 9.40 [1.75;19.70] | 236.50 [138.15;459.38] | < 0.001 |
PCT (ng/mL) | 0.06 [0.04;0.09] | 0.34 [0.26;0.88] | < 0.001 |
LDH (U/L) | 199.00 [170.50;275.50] | 341.50 [298.75;446.00] | 0.001 |
D-Dimer (µg/L) | 692.00 [336.50;1619.50] | 2392.00 [1525.00;2925.50] | 0.020 |
cfDNA (ng/mL) | 8.35 [4.91;12.94] | 24.62 [19.21;39.06] | < 0.001 |