Additional file 1: of Plasma myeloperoxidase-conjugated DNA level predicts outcomes and organ dysfunction in patients with septic shock

Figure S1. Determination of linearity of the MPO-DNA assay. The linear range of optical density (OD) in the MPO-DNA assay was determined with various sample volumes: A) 0, 6.25, 12.5, 25, 50, 75, and 100 μl; B) 0, 6.25, 12.5, 25, and 50 μl (n = 4 for each sample volume). (PPTX 64 kb

Additional file 2: of Plasma myeloperoxidase-conjugated DNA level predicts outcomes and organ dysfunction in patients with septic shock

Figure S2. Correlations of MPO-DNA and cf-DNA levels with organ failure parameters. Correlations of MPO-DNA and cf-DNA levels with the MAP (A), the P/F ratio (B), and the SOFA score (C) on day 1 after the diagnosis of septic shock. (PPTX 114 kb

Additional file 3: of Plasma myeloperoxidase-conjugated DNA level predicts outcomes and organ dysfunction in patients with septic shock

Figure S3. Correlations of MPO-DNA and cf-DNA levels with the platelet count and the DIC score. Correlations of MPO-DNA and cf-DNA levels with the platelet count (A) and the DIC score (B) on day 3 after the diagnosis of septic shock. (PPTX 76 kb

Baseline characteristics of the patients with septic shock and the controls.

<p>Baseline characteristics of the patients with septic shock and the controls.</p

Increased PD-1 Expression and Altered T Cell Repertoire Diversity Predict Mortality in Patients with Septic Shock: A Preliminary Study

<div><p>Sepsis causes impairment of innate and adaptive immunity by multiple mechanisms, including depletion of immune effector cells and T cell exhaustion. Although lymphocyte dysfunction is associated with increased mortality and potential reactivation of latent viral infection in patients with septic shock, the relation between viral reactivation and lymphocyte dysfunction is obscure. The objectives of this study were 1) to determine the relation of lymphocyte dysfunction to viral reactivation and mortality, and 2) to evaluate recovery of lymphocyte function during septic shock, including T cell receptor (TCR) diversity and the expression of programmed death 1 (PD-1). In 18 patients with septic shock and latent cytomegalovirus (CMV) infection, serial blood samples were obtained on days 1, 3, and 7 after the onset of shock, and immune cell subsets and receptor expression were characterized by flow cytometry. TCR diversity of peripheral blood mononuclear cells was analyzed by Multi-N-plex PCR, and CMV DNA was quantified using a real-time PCR kit. A decrease of TCR diversity and monocyte HLA-DR expression were observed in the early stage of septic shock, while CD4+ T cells displayed an increase of PD-1 expression. Significant lymphopenia persisted for at least 7 days following the onset of septic shock. Normalization of TCR diversity and PD-1 expression was observed by day 7, except in patients who died. CMV reactivation was detected in 3 of the 18 patients during the first week of their ICU stay and all 3 patients died. These changes are consistent with the early stage of immune cell exhaustion and indicate the importance of normal lymphocyte function for recovery from septic shock. Ongoing lymphocyte dysfunction is associated with CMV reactivation and dissemination, as well as with unfavorable outcomes.</p></div

Immune cell surface antigen expression, lymphocyte count, and TCR diversity in septic shock.

<p>TCR diversity (expressed as a percentage) was calculated as the ratio of the observed number of rearrangements to the theoretical number (A). Lymphocyte count (B), PD-1 expression by CD4+ T cells (C), and HLA-DR expression by CD14+ monocytes (D) measured on day 1, day 3, and day 7 after the diagnosis of septic shock compared with that in healthy volunteers (HV). Data are shown as box plot with medians (lines inside boxes), 25^th and 75^th quartiles (lines of boxes) and whiskers indicate the range. Any data not included between the whiskers were plotted as an outlier with small circle. **, p<0.01, *p<0.05 vs. healthy volunteers; ††, p<0.01, †, p<0.05 vs. day 1.</p

TCR diversity plotted versus PD-1 expression on quadrant charts for days 1, 3, and 7.

<p>Each chart is divided into four zones limited by the normal threshold of TCR diversity (78.3% based on lower quantile data from healthy volunteers) and the normal threshold of PD-1 expression (19.5% based on upper quantile data from healthy volunteers). Patients with CMV reactivation who died are indicated by closed triangles on day 7 and by open triangles on days 1 and 3. Group A: high PD-1 expression with low TCR diversity; group B: high PD-1 expression with normal TCR diversity; group C: normal PD-1 expression with low TCR diversity; group D: normal PD-1 expression with normal TCR diversity. Results are presented as individual values.</p

Comparison of CDR3 region of the TCR β family between dnTGFβRII and dnTGFβRII IL-6^−/− mice.

<p>The arrows indicate clonal expansion of specific Vβ. C57B6 mice were used as negative control. With this technique, if there is no detectable T cell expansion within a Vβ spectrum, a Gaussian distribution of CDR3 lengths is observed. In contrast, clonal expansions are observed as a perturbation of this Gaussian distribution.</p

IL-12p40 is required for 2OA-BSA-induced cholangitis.

<p>(A) H&amp;E staining of representative liver from WT, IL-12p40^−/− (p40^−/−), IL-23^−/− (p19^−/−) and IL-12p35^−/− (p35^−/−) mice 8 weeks after 2OA-BSA immunization. Portal inflammatory changes with interlobular bile duct damage (red arrow) were observed in the liver of WT, p19^−/− and p35^−/− mice but not p40^−/− mice; normal bile ducts in p40^−/− mice (blue arrow). (B) Portal inflammation and bile duct damage were examined in individual animals. The pathological score of portal inflammation and biliary cell damage were evaluated in WT (n = 17), p19^−/− (n = 14), p35^−/− (n = 3) and p40^−/− (n = 5) mice. *p&lt;0.05, **p&lt;0.01, ***p&lt;0.001. (C) Immunohistochemical analysis of the liver of WT mice and IL-23p19^−/− mice at 8 weeks after 2OA-BSA immunization using mAbs to CD4 and CD8.</p

Inflammatory cytokine production in mice immunized with 2OA-BSA.

<p>(A) Serum levels of IL-17A, IFN-γ, TNF-α and IL-6 in WT, IL-17A^−/−, IL-17F^−/− and IL-22^−/− mice, respectively, at 2 weeks following 2OA-BSA immunization. (B) Production of IFN-γ in supernatant fluids of cultured splenic and hepatic MNCs (n = 4) with anti-CD3/CD28 mAbs at 3 days. (C) The level of inflammatory cytokines in extracted liver protein from WT, IL-17A^−/−, IL-17F^−/− and IL-22^−/− mice, respectively. Each group n = 8. *p&lt;0.05, **p&lt;0.01, ***p&lt;0.001.</p