Supplemental Digital Content
Multilateral functional alterations of human neutrophils in sepsis:
from the point of diagnosis to the seventh day
SHOCK
Akira Endo, M.D., Miko Okamura, M.Sc., Shunsuke Yoshikawa, M.D., Yasuhiro Otomo, M.D., Ph.D., Tomohiro Morio, M.D., Ph.D.
Correspondence:
Tomohiro Morio, M.D., Ph.D.
Department of Pediatrics and Developmental Biology, Graduate School of Tokyo Medical and Dental University
E-mail: [email protected]
eTable 1. Demographic and clinical characteristics of each patient.
Case Age Sex
Site of Infection
Causative microorganism
Blood culture
Septic shock
White blood cell count
SOFA score
APACHE II score
Outcome at discharge
1 95 F Abdomen Undetectable Negative No 11300 10 12 Died
2 77 M Soft tissue GPC Positive No 12800 6 11 Survived
3 71 M Abdomen Mixed Negative No 8000 0 11 Survived
4 74 F Lung GPC Positive No 20300 4 27 Survived
5 44 F Urinary tract GNR Positive Yes 9200 4 16 Survived
6 72 M Soft tissue GPC Negative No 8600 11 20 Survived
7 65 F Urinary tract GNR Positive No 3300 2 12 Survived
8 48 F Soft tissue Mixed Positive Yes 9700 2 21 Survived
9 82 F Abdomen GNR Positive Yes 2500 4 12 Survived
10 85 F Abdomen Mixed Positive Yes 21300 9 25 Survived
11 51 F Abdomen Undetectable Negative No 28500 8 19 Survived
12 89 F Urinary tract GNR Positive No 11600 0 12 Survived
13 69 F Abdomen Mixed Positive Yes 2100 5 21 Survived
14 86 M Abdomen Undetectable Negative Yes 21700 1 14 Survived
15 45 M Soft tissue GPC Positive Yes 2100 15 32 Died
16 87 F Lung Mixed Positive Yes 1700 11 25 Died
17 73 F Abdomen Undetectable Negative Yes 2300 4 14 Survived
18 64 F Abdomen Undetectable Negative No 700 5 12 Survived
19 63 M Abdomen Mixed Negative No 19800 7 8 Survived
20 69 M Lung GPC Negative Yes 19200 9 25 Survived
21 85 M Lung GNR Negative No 3400 11 30 Died
23 70 M Lung GNR Positive Yes 500 18 39 Died
24 77 M Lung GNR Negative No 9400 5 10 Survived
25 90 M Abdomen GNR Negative No 2100 3 22 Survived
26 88 F Abdomen Undetectable Negative Yes 12200 10 35 Died
27 66 F Abdomen GNR Negative No 5700 1 8 Survived
28 90 M Abdomen Mixed Positive No 1000 2 13 Survived
29 68 M Abdomen Mixed Negative No 3900 5 14 Survived
30 72 M Blood stream GPC Positive Yes 19800 9 23 Died
31 40 M Abdomen Mixed Negative No 10500 4 6 Survived
32 68 M Unclear Undetectable Negative No 1800 10 16 Survived
33 87 F Lung GPC Negative Yes 12700 7 19 Survived
34 43 M Abdomen Mixed Negative No 20200 1 3 Survived
35 47 M Abdomen Mixed Negative No 11800 4 6 Survived
36 39 M Abdomen Mixed Negative No 6700 2 3 Survived
37 83 M Abdomen Mixed Negative Yes 1900 4 21 Survived
38 89 F Abdomen GNR Positive Yes 17900 7 23 Survived
39 71 M Lung GNR Negative Yes 8200 5 16 Survived
40 83 F Abdomen Mixed Positive Yes 31800 17 21 Survived
41 87 F Abdomen Mixed Positive No 2800 3 13 Survived
42 67 M Abdomen Undetectable Negative Yes 34000 11 27 Survived
43 70 F Abdomen GNR Positive No 14600 2 10 Survived
44 68 M Blood stream GPC Positive Yes 6400 5 26 Died
45 79 M Urinary tract GNR Positive Yes 29200 4 19 Survived
46 79 F Abdomen Mixed Negative No 22000 0 13 Survived
47 75 M Soft tissue GPC Negative Yes 23100 15 41 Died
48 75 F Abdomen GNR Negative No 1600 3 19 Survived
49 77 M Soft tissue GPC Negative No 13100 2 12 Survived
50 82 F Abdomen Mixed Negative No 3800 2 23 Survived
51 81 M Lung GNR Negative No 20900 5 27 Survived
52 88 F Urinary tract GNR Positive No 18900 3 29 Survived
53 53 M Lung GPC Negative No 7500 4 11 Survived
54 32 M Abdomen GNR Negative No 24600 2 5 Survived
55 90 F Lung Mixed Positive Yes 19300 10 30 Survived
56 75 M Soft tissue Undetectable Negative No 14900 1 12 Survived
57 82 F Abdomen Mixed Negative No 600 4 14 Survived
58 56 M Lung GPC Positive No 18900 3 10 Survived
59 74 M Lung Undetectable Negative Yes 12900 3 20 Survived
Abbreviations: F, female; M, male; SOFA, Sequential Organ Failure Assessment; APACHE II, Acute Physiology and Chronic Health Evaluation II;
GPC, gram positive cocci; GNR, gram negative rod
eTable 2. Summary of performed treatments to the patients during study period.
Performed treatments Before day 0 Between day0 to day3 Between day3 to day7
Surgical interventions, n (%) 0 / 59 (0) 18 / 30 (60.0) 0 / 24 (0)
Corticosteroids, n (%) 0 / 59 (0) 1 / 30 (3.4) 1 / 24 (4.2)
Immunoglobrin, n (%) 0 / 59 (0) 2 / 30 (6.7) 0 / 24 (0)
chathechoramines, n (%) 0 / 59 (0) 14 / 30 (46.7) 4/ 24 (16.7)
Transfusion, n (%) 0 / 59 (0) 7 / 30 (23.3) 1 / 24 (4.2)
Antibiotics
Carbapenem, n (%) 3 / 59 (5.1) 11 / 30 (36.7) 5 / 24 (20.8)
Cephalosporin, n (%) 4 / 59 (6.8) 8 / 30 (26.7) 9 / 24 (37.5)
Tazobactam/piperacillin, n (%) 3 / 59 (5.1) 6 / 30 (20.0) 5 / 24 (20.8) Ampicillin/sulbactam, n (%) 4 / 59 (6.8) 1 / 30 (3.4) 3 / 24 (12.5)
Others, n (%) 0 / 59 (0) 4 / 30 (13.3) 1 / 24 (0)
Ventilator, n (%) 0 / 59 (0) 12 / 30 (40.0) 7 / 24 (29.2)
Hemofiltration, n (%)
0 / 59 (0) 1 / 30 (3.4) 0 / 24 (0)
Polymyxin B hemoperfusion, n (%) 0 / 59 (0) 3 / 30 (10.0) 0 / 24 (0)
eFigure 1. Comparison of serum cytokine concentrations between survivors and non-survivors.
The sample size in each group was described in parentheses. The Mann–Whitney U-test was used for comparison
between survivors and non-survivors. Abbreviations: IL, interleukin; TNF, tumor necrosis factor; IFN, interferon
eFigure 2. Correlations between the initial serum cytokine concentrations and the clinical scores
The solid line shows the regression line. Patients who survived are presented in blue, and patients who died are
presented in red. Abbreviations: APACHE II, Acute Physiology and Chronic Health Evaluation II; SOFA,
Sequential Organ Failure Assessment; IL, interleukin; TNF, tumor necrosis factor, R, correlation coefficient
eFigure 3. Correlation between serum IL-6 level and IL-10 level in each individual
The solid line shows the regression line. Patients who survived are presented in blue, and patients who died are
presented in red. Abbreviations: IL, interleukin; R, correlation coefficient
eFigure 4. Comparison of reactive oxygen species production between survivors and non-survivors.
The sample size in each group was described in parentheses. The Mann–Whitney U-test was used for comparison
between survivors and non-survivors. Abbreviations: ROS, reactive oxygen species; RLU, relative light unit; fMLP,
N-Formylmethionyl-leucyl-phenylalanine; PMA, 12-myristate 13-acetate
eFigure 5. Correlations between the neutrophil priming state and serum cytokine concentrations
The solid line shows the regression line. Patients who survived are presented in blue, and patients who died are
presented in red. Abbreviations: IL, interleukin; TNF, tumor necrosis factor; fMLP, N-Formylmethionyl-leucyl-
phenylalanine; ROS, reactive oxygen species; RLU, relative light unit; R, correlation coefficient
eFigure 6. Reactive oxygen species release pattern in a representative patient and a healthy volunteer.
Abbreviations: ROS, reactive oxygen species; fMLP, N-Formylmethionyl-leucyl-phenylalanine; PMA, 12-myristate
13-acetate
eFigure 7. Correlation between the expression of flavocytochrome b
558and neutrophil priming state
The solid line shows the regression line. Patients who survived are presented in blue, and patients who died are
presented in red. Abbreviations: fMLP, N-Formylmethionyl-leucyl-phenylalanine; ROS, reactive oxygen species;
RLU, relative light unit; MFI, mean fluorescence intensity; R, correlation coefficient
eFigure 8. Correlation between the expression of CXCR2 and the concentrations of serum cytokine
The solid line shows the regression line. Patients who survived are presented in blue, and patients who died are
presented in red. Abbreviations: IL, interleukin; TNF, tumor necrosis factor; CXCR2, CXC chemokine receptor 2;
MFI, mean fluorescence intensity; R, correlation coefficient
eFigure 9. Cluster distribution and histogram of Annexin V and 7-aminoactinomycin staining.
Apoptosis assay; the histogram of 7-AAD showed a bimodal distribution and the histogram of Annexin V showed a
monomodal distribution. Abbreviation: 7-AAD, 7-aminoactinomycin
