Upregulation of the pro-apoptotic genes BID and FAS in septic shock patients

Introduction: Lymphocyte apoptosis has been suggested to play a central role in sepsis pathophysiology, and studies in animal models demonstrated that blocking this pathway improves outcome. However, no routine biomarkers of apoptosis are so far available in patients. Thus, the aim of our study was to assess the different biomarkers of apoptosis putatively usable on a routine basis in septic shock.Methods: Thirteen septic shock patients (sampled twice between days 1 to 2 and days 3 to 5 after diagnosis of shock) and 15 sex-matched and age-matched healthy controls were prospectively enrolled. Apoptosis was measured in lymphocyte subpopulations using flow cytometry (Annexin-V binding, activated caspase-3 and Bcl-2 expressions). Representative pro-apoptotic and anti-apoptotic gene expressions were assessed by quantitative reverse-transcription PCR. Monocyte HLA-DR expression and lymphocyte subpopulation cell counts were measured as markers of sepsis-induced immune dysfunctions. To test for statistical significance, the Mann-Whitney U test was used with correction by the number of tests performed.Results: Flow cytometric measurements of apoptosis in septic shock patients showed an increased Annexin-V binding on CD4+ T cells and an increased active caspase-3 expression on B cells only at days 3 to 5 (sixfold change and twofold change, respectively). Gene expression analysis showed an increased BCL-XL mRNA and anupregulation of the pro-apoptotic genes BID and FAS in septic shock patients (10-fold change and fivefold change, respectively) compared with healthy controls.Conclusions: The present study highlights the difficulties encountered in monitoring apoptosis on a routine basis in septic patients, whereas in the same sampling conditions and on the same patients, HLA-DR expression and lymphocyte subpopulation cell counts showed characteristics described in the literature. However, pro-apoptotic genes BID and FAS appear to constitute promising apoptosis markers in our hands

Early Interleukin-6 and Slope of Monocyte Human Leukocyte Antigen-DR: A Powerful Association to Predict the Development of Sepsis after Major Trauma

OBJECTIVE: Major trauma is characterized by a pro-inflammatory response, followed by an immunosuppression. Recently, in trauma patients, the lack of recovery of monocyte Human Leukocyte Antigen DR (mHLA-DR, a biomarker of ICU-acquired immunosuppression) between days 1-2 and days 3-4 has been demonstrated to be independently associated with sepsis development. The main objective of this study was to determine whether early measurements of IL-6 (interleukin-6) and IL-10 plasma concentrations (as markers of initial severity) could improve, in association with mHLA-DR recovery, the prediction of sepsis occurrence in severe trauma patients. DESIGN: Prospective observational study over 24 months in a Trauma ICU at university hospital. PATIENTS: Trauma patients with an ISS over 25 and age over 18 were included. MEASUREMENTS AND MAIN RESULTS: mHLA-DR was assessed by flow cytometry, IL-6 and IL-10 concentrations by ELISA. 100 consecutive severely injured patients were monitored (mean ISS 37±10). 37 patients developed sepsis. IL-6 concentrations and slope of mHLA-DR expression between days 1-2 and days 3-4 were significantly different between septic and non-septic patients. IL-10 was not detectable in most patients. After adjustment for usual clinical confounders, when assessed as a pair, multivariate logistic regression analysis revealed that a slope of mHLA-DR expression (days 3-4/days 1-2)≤1.1 and a IL-6 concentration ≥ 67.1 pg/ml remained highly associated with the development of sepsis (adjusted OR 18.4, 95% CI 4.9; 69.4, p = .00002). CONCLUSIONS: After multivariate regression logistic analysis, when assessed as a pair, a high IL-6 concentration and a persistent mHLA-DR decreased expression were found to be in relation with the development of sepsis with the best predictive value. This study underlines the usefulness of daily monitoring of immune function to identify trauma patients at a high risk of infection

Programmed death-1 levels correlate with increased mortality, nosocomial infection and immune dysfunctions in septic shock patients

International audienceINTRODUCTION: Septic shock remains a major health care problem worldwide. Sepsis-induced immune alterations are thought to play a major role in patients' mortality and susceptibility to nosocomial infections. Programmed death-1 (PD-1) receptor system constitutes a newly described immunoregulatory pathway that negatively controls immune responses. It has recently been shown that PD-1 knock-out mice exhibited a lower mortality in response to experimental sepsis. The objective of the present study was to investigate PD-1-related molecule expressions in septic shock patients. METHODS: This prospective and observational study included 64 septic shock patients, 13 trauma patients and 49 healthy individuals. PD-1-related-molecule expressions were measured by flow cytometry on circulating leukocytes. Plasmatic interleukin (IL)-10 concentration as well as ex vivo mitogen-induced lymphocyte proliferation were assessed. RESULTS: We observed that septic shock patients displayed increased PD-1, PD-Ligand1 (PD-L1) and PD-L2 monocyte expressions and enhanced PD-1 and PD-L1 CD4+ T lymphocyte expressions at day 1-2 and 3-5 after the onset of shock in comparison with patients with trauma and healthy volunteers. Importantly, increased expressions were associated with increased occurrence of secondary nosocomial infections and mortality after septic shock as well as with decreased mitogen-induced lymphocyte proliferation and increased circulating IL-10 concentration. CONCLUSIONS: These findings indicate that PD-1-related molecules may constitute a novel immunoregulatory system involved in sepsis-induced immune alterations. Results should be confirmed in a larger cohort of patients. This may offer innovative therapeutic perspectives on the treatment of this hitherto deadly disease

Biomarkers in severe sepsis : toward new individualized therapeutic strategies

En dépit de nombreux essais thérapeutiques, les syndromes septiques sont la première cause de mortalité en service de soins intensifs. La population septique étant très hétérogène, une meilleure caractérisation des patients serait essentielle afin de mieux individualiser et cibler les thérapeutiques potentiellement bénéfiques. Une approche multiparamétrique de l’utilisation des biomarqueurs est une alternative qui viserait à appréhender la situation de manière plus globale. Notre travail s’inscrit dans ce contexte au travers de l’étude plus spécifique de la défaillance des systèmes cardio-vasculaire et immunitaire. Au-delà de la confirmation de l’intérêt des biomarqueurs présentement étudiés (prohormones cardio-vasculaires et PD-1) dans la prédiction de la mortalité et du risque d’infections nosocomiales, nos résultats apportent des éléments nouveaux. Nous avons montré que (1) la sur-expression des molécules PD-1 est associée à l’énergie leucocytaire, (2) un même biomarqueur peut apporter une information différente au cours du temps, (3) l’information apportée par l’analyse simultanée de deux biomarqueurs est supérieure à celle de la somme de leurs valeurs individuelles, et (4) l’expression dynamique d’un biomarqueur est meilleure que son expression à un temps donné. Au total, notre travail illustre l’intérêt potentiel d’un panel de biomarqueurs pour mieux appréhender la complexité des états septiques et leur rapide évolution. Il reste néanmoins à développer des outils biostatistiques capables de donner au clinicien une vision globale en temps réel des processus en cours. Cela constituera une étape clé pour mieux stratifier et cibler les prochains essais cliniques dans le domaine.Septic syndromes remain the leading cause of death in the intensive care units despite numerous clinical trials. Septic patients constitute a very heterogeneous population. Therefore improved characterisation of patients in order to better target and personalize potential new therapeutics is highly desirable. A multiparametric biomarker-based approach could be an attractive alternative to obtain a global view of the pathophysiologic situation. In this context, we worked specifically on cardio-vascular and immune dysfunctions. We first confirmed the predictive value of biomarkers for mortality or nosocomial infections, and showed new elements. We observed that (1) PD-1 overexpression is associated with leukocyte anergy, (2) one biomarker could give different information over time, (3) information provided by the association of two biomarkers is more interesting than the addition of their individual values, and (4) dynamic expression of one biomarker is more informative than its expression at a given time point. Finally, our results illustrate the potential interest of biomarker panels to improve our understanding of the septic syndrome complexity and to reflect their rapid evolution. Consequently, next step will depend on our capacity to develop biostatistic tools that enable clinicians to get, in real time, a global view of the process over time. This key step is likely necessary to decrease the heterogeneity of septic patient population in order to better stratify and target next clinical trials in the field

Biomarqueurs des états septiques sévères : vers de nouvelles stratégies thérapeutiques individualisées

Septic syndromes remain the leading cause of death in the intensive care units despite numerous clinical trials. Septic patients constitute a very heterogeneous population. Therefore improved characterisation of patients in order to better target and personalize potential new therapeutics is highly desirable. A multiparametric biomarker-based approach could be an attractive alternative to obtain a global view of the pathophysiologic situation. In this context, we worked specifically on cardio-vascular and immune dysfunctions. We first confirmed the predictive value of biomarkers for mortality or nosocomial infections, and showed new elements. We observed that (1) PD-1 overexpression is associated with leukocyte anergy, (2) one biomarker could give different information over time, (3) information provided by the association of two biomarkers is more interesting than the addition of their individual values, and (4) dynamic expression of one biomarker is more informative than its expression at a given time point. Finally, our results illustrate the potential interest of biomarker panels to improve our understanding of the septic syndrome complexity and to reflect their rapid evolution. Consequently, next step will depend on our capacity to develop biostatistic tools that enable clinicians to get, in real time, a global view of the process over time. This key step is likely necessary to decrease the heterogeneity of septic patient population in order to better stratify and target next clinical trials in the field.En dépit de nombreux essais thérapeutiques, les syndromes septiques sont la première cause de mortalité en service de soins intensifs. La population septique étant très hétérogène, une meilleure caractérisation des patients serait essentielle afin de mieux individualiser et cibler les thérapeutiques potentiellement bénéfiques. Une approche multiparamétrique de l’utilisation des biomarqueurs est une alternative qui viserait à appréhender la situation de manière plus globale. Notre travail s’inscrit dans ce contexte au travers de l’étude plus spécifique de la défaillance des systèmes cardio-vasculaire et immunitaire. Au-delà de la confirmation de l’intérêt des biomarqueurs présentement étudiés (prohormones cardio-vasculaires et PD-1) dans la prédiction de la mortalité et du risque d’infections nosocomiales, nos résultats apportent des éléments nouveaux. Nous avons montré que (1) la sur-expression des molécules PD-1 est associée à l’énergie leucocytaire, (2) un même biomarqueur peut apporter une information différente au cours du temps, (3) l’information apportée par l’analyse simultanée de deux biomarqueurs est supérieure à celle de la somme de leurs valeurs individuelles, et (4) l’expression dynamique d’un biomarqueur est meilleure que son expression à un temps donné. Au total, notre travail illustre l’intérêt potentiel d’un panel de biomarqueurs pour mieux appréhender la complexité des états septiques et leur rapide évolution. Il reste néanmoins à développer des outils biostatistiques capables de donner au clinicien une vision globale en temps réel des processus en cours. Cela constituera une étape clé pour mieux stratifier et cibler les prochains essais cliniques dans le domaine

Evaluation de quatre précurseurs d'hormones vasoactives comme marqueurs prédictifs de la mortalité au cours du choc septique

LYON1-BU Santé (693882101) / SudocSudocFranceF

Physiopathologie et conséquences de l'immunodépression acquise en réanimation

International audienc