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SUPPLEMENTARY MATERIALS Supplementary Table 1: Antibodies used in this study Antibody. Manufacturer. (FITC) conjugated CD36 monoclonal antibody (mAb) (CD36-FITC, clone FA6.152). Immunotech. phycoerythrin (PE) conjugated CD2 mAb (CD2-PE, clone 39C1.5). (Marseille, France).. PE conjugated CD294 mAb (CD294-PE, clone BM16) PE Texas Red (PE-TR so called ECD) conjugated CD19 mAb (CD19-ECD, clone J3-119) PE-cyanine 5 (Cy5) conjugated CD16 mAb (CD16-PC5, clone 3G8) PE-cyanine 7 (Cy7) conjugated CD45 mAb (CD45-PC7, clone J33) FITC conjugated CD14 mAb (CD14-FITC,clone RMO52) ECD conjugated CD45 mAb (CD45-ECD,clone j.33) PC5 conjugated CD11b mAb (CD11b-PC5, clone Bear1) PC7 conjugated CD16 mAb (CD16-PC7, clone 3G8) PE conjugated CD11c mAb (CD11c-PE, Clone BU15) FITC conjugated CD64 mAb (CD64-FITC, clone 22) PE conjugated CD10 mAb (CD10-PE,clone ALB1) PC5 conjugated CD24 mAb (CD24-PC5, clone ALB9) PC7 conjugated CD34 mAb (CD34-PC7, clone 581) PE conjugated CD123 mAb (CD123-PE,clone SSDCLY107D2) PC5 conjugated CD138 mAb (CD138-PC5, clone BB4) PC7 conjugated CD4 mAb (CD4-PC7, clone SFCI12T4D11) FITC conjugated CD4 mAb (CD4-FITC, 13B8.2) PE conjugated DR mAb (DR, Immu-357) ECD conjugated CD8 mAb (CD8-ECD, clone SFC21Thy 2D3) PC5.5 conjugated CD38 mAb (CD38 PC5.5, clone LS198-4-3) PC7 conjugated CD25 mAb (CD25-PC7, clone B1.49.9) Allophycocyanin (APC) conjugated CD56 mAb (CD56-APC, clone N901(NKH-1) APC Alexa Fluor (A) 700 conjugated CD127 mAb (CD127-APC-A700, clone R34.34) APC-A750 conjugated CD3 mAb (CD3-APC-A750, clone UCHT1) Pacific blue (PB) conjugated CD16 mAb (CD16-PB, clone 3G8) Krome Orange (KO) conjugated CD45 mAb (CD45-KO, clone J.33) FITC conjugated CD116 mAb (CD116-FITC, clone M5D12),. Becton Dickinson. 1. Supplementary Table 2 : Presumed deduced cell types from the gating properties of each cell studied Variable median [Q1;Q3]* Lymphocytes G/L. Combination. Gating properties. (iv). SSC low,CD45 high. (iv). SSC low,CD45 high,CD19+. T-lymphocytes, CD3pos, G/L. (iv). SSC low,CD45 high,CD3+. T-lymphocytes, CD4pos, G/L. (iv). SSC low,CD45 high,CD3+,CD4+. T-lymphocytes, CD8pos, G/L. (iv). SSC low,CD45 high,CD3+,CD8+. NK lymphocytes, G/L. (iv). SSC low,CD45 high,CD3-,CD16+ or CD56+. NK-like lymphocytes, G/L. (iv). SSC low,CD45 high,CD3-,CD16+ or CD56+. Activated CD4pos lymphocytes, G/L. (iv). SSC low,CD45 high,CD3+,CD4+, DR+ or CD25+ or CD38+. Activated CD8pos lymphocytes, G/L. (iv). SSC low,CD45 high,CD3+,CD8+,. T reg, G/L. (iv). SSC low,CD45 high,CD3+,CD25+, CD127 dim. Granulocytes G/L :. (iii). SSC high/CD45int. - increased CD64 expression, %**. (iii). SSC high/CD45int/CD64+. - decreased CD16 expression, %**. (ii). SSC high/CD45int/CD14-/CD11b-CD16dim. - decreased CD10 expression, % **. (iii). SSC high/CD45int/CD10dim. - increased CD24 expression, %**. (iii). SSC high/CD45int/CD24+. Hematogones, G/L. (iii). SSC low/CD45int/cd16-/CD64-/not CD45 high/CD10+. DC1, G/L. (v). CD45int/not SShigh/CD4int/not CD123-/CD116+. DC2, G/L. (v). CD45int/not SShigh/CD4int/not CD123+. CD34+, G/L. (iii). SSC low/CD45int/CD34+. Plasmocytes G/L. (v). CD45+/CD138+. Promonocytes, G/L. (ii). SSCInt/CD45 high/CD14Int/CD11bInt. Monocytes, G/L. (ii). SSCInt/CD45 high/CD14+. increased CD16 expression, %. (ii). SSCInt/CD45 high/CD14+/CD16+. increased DRneg monocytes %. (iv). CD45int/not SShigh/CD4+. B-lymphocytes , CD19pos, G/L. DR+ or CD25+ or CD38+. (i) (ii) (iii) (iv) (v). CD36-FITC / CD2-PE + CD294-PE /CD19-ECD / CD16-PC5 /CD45-PC7, CD14-FITC / CD11c-PE / CD45-ECD / CD11b-PC5 /CD16-PC7, CD64-FITC / CD10-PE / CD45-ECD / CD24-PC5 / CD34-PC7 CD4-FITC /DR-PE / CD8-ECD/ CD38-PC5/ CD25-PC7 /CD56-APC / CD127-A700 /CD3A750 /CD16-PB / CD45-KO CD116-FITC / CD123-PE / CD45-ECD / CD138-PC5 / CD4-PC7.. 2. Supplementary Table 3: comparison between controls and patients with sepsis at diagnosis; * Wilcoxon’s test Parameters [Q1;Q3]*. GB, G/L Haemoglobin, g/dL Platelet, G/L Lymphocytes G/L. Controls (non infected) (n=20). Patients (n=177). p-value**. 6.7 [5.3;7.6]. 14.05 [9.77.18.9]. <0.001. 14 [12.9 ; 15]. 13 [11.7;14.2]. 0.001. 243.5 [205.75;286.75]. 258.5[186.75 ;319.25]. 0.780. 1.82 [1.52 ; 2.2]. 0.865 [0.648 ; 1.403]. < 0.001. B-lymphocytes, CD19pos, G/L. 0.199 [0.115 ; 0.268]. 0.123 [0.083 ; 0.234]. 0.02. T-lymphocytes, CD3pos, G/L. 1.278[1.036 ; 1.683]. 0.541 [0.348 ; 0.93]. <0.001. T-lymphocytes, CD4pos, G/L. 0.769 [0.554 ; 0.918]. 0.322 [0.188 ; 0.543]. <0.001. T-lymphocytes, CD8pos, G/L. 0.413[0.296 ; 0.578]. 0.164 [0.079 ;0.301]. <0.001. NK-lymphocytes, G/L. 0.259 [0.152 ; 0.374]. 0.154 [0.079 ; 0.263]. <0.001. NK-like lymphocytes, G/L. 0.031[0.019;0.068]]. 0.02 [0.007;0.047]. 0.002. Activated CD4pos lymphocytes, G/L. 0.318 [0.22;0.478]. 0.136 [0.075; 0.204]. <0.001. Activated CD8pos lymphocytes, G/L. 0.11 [0.076;0.146]. 0.05 [0.027 ;0.092]. <0.001. T reg, G/L. 0.046 [0.033; 0.059]. 0.019 [0.011;0.032]. <0.001. Granulocytes G/L :. <0.001. 3.672 [2.863 ; 4.636]. 11.657 [8.122 ; 16.832]. - increased CD64 expression, %*. 0.2 [0.15 ; 0.25]. 14.045 [1.198 ; 54.683]. - decreased CD16 expression, %*. 0.372 [0.267 ; 0.631]. 6.33 [1.833 ; 22]. <0.001. - decreased CD10 expression, % *. 0.275 [0.02 ;0.862]. 68.435 [32.62 ; 93.44]. <0.001. - increased CD24 expression, %*. 2.15 [0.045 ;0.815]. 19.351 [0.75. 27.2]. Hématogones, G/L. 0.005 [0.003;0.011]. 0.002 [0;0.004]. DC1, G/L. 0.002 [0;0.0007]. 0.001 [0;0.0008]. 0.726. DC2, G/L. 0.009 [0.006;0.011]. 0.001 [0;0.004]. <0.001. CD34+, G/L. 0.002 [0.001;0.003]. 0.002 [0.001;0.003]. 0.955. 0.001 [0;0.001]. 0.001 [0.001;0.002]. 0.141. Promonocytes, G/L. 0.014 [0.011 ; 0.018]. 0.008 [0.004 ; 0.018]. <0.001. Monocytes, G/L. 0.528 [0.406 ; 0.72]. 0.811 [0.434 ; 1.213]. 0.001. increased CD16 expression, %***. 0.057 [0.041 ; 0.08]. 0.076 [0.037 ; 0.125]. 0.073. increased DRneg monocytes %***. 7.85 [5.47 ; 16.69]. 22.51 [10.27 ; 38.23]. <0.001. Plasmocytes G/L. <0.001. **The threshold has been established from a control group and the percentage is given on granulocyte population among granulocytes; *** The threshold has been established from a control group and the percentage is given on granulocyte population among granulocytes;. 3. Supplementary Table 4: Threshold and AUC from comparison between controls and patients with sepsis at diagnosis Parameters. Threshold*. AUC. 95% Confidence interval**. Sensitivity. Specificity. p-value*** (Area=0.5). GB, G/L. >10.5. 0.879. 0.829-0.919. 100. 73.4. <0.0001. Haemoglobin, g/dL. ≤12.7. 0.655. 0.589-0.717. 43.5. 85.1. 0.0001. -. 0.522. 0.454-0.590. Lymphocytes G/L. ≤1.258. 0.835. 0.781-0.881. 71.2. 89.8. <0.0001. B-lymphocytes, CD19pos, G/L. ≤0.1354. 0.602. 0.535-0.666. 56.5. 69.4. 0.0220. T-lymphocytes, CD3pos, G/L. ≤0.9323. 0.850. 0.796-0.894. 76. 88. <0.0001. T-lymphocytes, CD4pos, G/L. ≤0.419. 0.817. 0.760-0.865. 63.3. 91.8. <0.0001. T-lymphocytes, CD8pos, G/L. ≤0.2693. 0.808. 0.751-0.857. 69.5. 81.6. <0.0001. NK-lymphocytes, G/L. ≤0.1078. 0.699. 0.635-0.758. 39. 98. <0.0001. NK-like lymphocytes, G/L. ≤0.0161. 0.646. 0.579-0.708. 46.3. 85.7. 0.0002. Activated CD4pos lymphocytes, G/L. ≤0.2157. 0.841. 0.787-0.886. 79.7. 79.6. <0.0001. Activated CD8pos lymphocytes, G/L. ≤0.0646. 0.784. 0.725-0.836. 61. 89.8. <0.0001. T reg, G/L. ≤0.0282. 0.833. 0.778-0.879. 66.7. 85.7. <0.0001. Granulocytes G/L :. >6.29. 0.932. 0.891-0.961. 83.6. 98. <0.0001. - increased CD64 expression, %**. >0.55. 0.924. 0.882-0955. 84. 98. <0.0001. - decreased CD16 expression, %**. >0.8357. 0.919. 0.875-0.951. 86. 86. <0.0001. - decreased CD10 expression, % **. >6.84. 0.984. 0.971-0.999. 96. 96. <0.0001. - increased CD24 expression, %**. >1.34. 0.841. 0.786-0.886. 70.6. 83.3. <0.0001. Hématogones, G/L. ≤0.0034. 0.760. 0.699-0.814. 74.6. 67.3. <0.0001. 0.523. 0.456-0.591. 0.904. 0.857-0.939. 0.502. 0.435-0.569. -. -. 0.9611. -. -. 0.0945. Platelet, G/L. DC1, G/L DC2, G/L CD34+, G/L. ≤0.0047 -. -. -. 81. 94. 0.5791. 0.6356 <0.0001. Plasmocytes G/L. >0.0012. 0.568. 0.500-0.633. Promonocytes, G/L. ≤0.0093. 0.662. 0.595-0.724. 59.6. 83.7. <0.0001. Monocytes, G/L. >0.7656. 0.658. 0.592-0.721. 50.9. 85.7. <0.0001. increased CD16 expression, %. 0.549. 0.479-0.615. increased DRneg monocytes %***. 0.757. 0.0.695-0.812. 71.8. 74. 0.2118 <0.001. * criterion corresponding with highest Younden Index; ** Binomial exact ***ROC. 4. Supplementary Table 5: Threshold and AUC from comparison between stable or improvement group and deteriorating group Parameters. Threshold * ≤7.2. AUC. 95% Confidence interval**. Sensitivity. Specificity. p-value***. 0.668. 0.593 to 0.737. 41.3. 92.4. 0.0013. Haemoglobin, g/dL. -. 0.516. 0.440 to 0.591. -. -. 0.7698. Platelet, G/L. -. 0.558. 0.481 to 0.633. -. -. 0.2941. ≤1.09. 0.601. 0.524 to 0.673. -. 0.538. 0.461 to 0.613. GB, G/L. Lymphocytes G/L B-lymphocytes, CD19pos, G/L. 76.1. 42.7 -. 0.0389 -. 0.4801. T-lymphocytes, CD3pos, G/L. ≤0.5567. 0.616. 0.540 to 0.688. 69.6. 53.4. 0.0174. T-lymphocytes, CD4pos, G/L. ≤ 0.3874. 0.635. 0.559 to 0.706. 73.9. 49.6. 0.0039. T-lymphocytes, CD8pos, G/L. ≤0.097. 0.598. 0.522 to 0.671. -. -. 0.0587. NK-lymphocytes, G/L. ≤0.0818. 0.574. 0.498 to 0.648. -. -. 0.1249. NK-like lymphocytes, G/L. ≤ 0.0049. 0.536. 0.459 to 0.611. -. -. 0.531. Activated CD4pos lymphocytes, G/L. ≤ 0.2224. 0.609. 0.553 to 0.682. Activated CD8pos lymphocytes, G/L. ≤ 0.0229. 0.597. 0.521 to 0.670. T reg, G/L. ≤ 0.0161. 0.647. 0.572 to 0.717. 56.5. 67.2. 0.0014. Granulocytes G/L :. ≤ 6.5071. 0.660. 0.586 to 0.730. 41.3. 91.6. 0.002. - increased CD64 expression, %**. >1.09. 0.572. 0.495 to 0.645. - decreased CD16 expression, %**. >15. 0.813. 0.748 to 0.868. 69.6. 83.2. <0.0001. - decreased CD10 expression, % **. >92. 0.713. 0.640 to 0.718. 56.5. 80.9. <0.0001. - increased CD24 expression, %**. >32. 0.610. 0.534 to 0.683. 37. 84.4. 0.0254. Hématogones, G/L. ≤ 0.0011. 0.626. 0.550 to 0.697. 52. 71. 0.0074. DC1, G/L. >0.0008. 0.557. 0.480 to 0.632. DC2, G/L. ≤ 0.0013. 0.652. 0.577 to 0.722. 95.7. 27.4 -. 0.021 -. 0.0674. 0.1156. 73.9. 56.6. 0.2519 0.0009. CD34+, G/L. -. 0.513. 0.436 to 0.589. -. -. 0.7893. Plasmocytes G/L. -. 0.537. 0.461 to 0.612. -. -. 0.4470. Promonocytes, G/L. -. 0.595. 0.517 to 0.669. -. -. 0.0785. Monocytes, G/L. ≤0.4038. 0.665. 0.589 to 0.735. increased CD16 expression, %. >20. 0.563. 0.486 to 0.639. increased DRneg monocytes %. >30. 0.678. 0.602 to 0.748. 44.4. 87.3. 0.0015 0.2109. 56. 76. 0.0004. * criterion corresponding with highest Younden Index; **Wilcoxon’s test *** ROC. 5.