Christelle Marie

Ex vivo T-lymphocyte derived cytokine production in SIRS patients is influenced by experimental procedures.

ABSTRACT Ex vivo production of interleukin‐2 (IL‐2), IL‐4, IL‐5, and IL‐10 by peripheral blood mononuclear cells (PBMC) was studied in 13 septic patients with infectious systemic inflammatory response syndrome (SIRS) and 13 patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) (noninfectious SIRS). We have investigated the levels of cytokines after activation by either concanavalin A (ConA), phytohemagglutinin (PHA), or anti‐CD3 antibodies. In whole blood assays, ConA‐induced IL‐10 was significantly reduced in both groups of patients compared with healthy controls. In sepsis patients, IL‐2, IL‐5, and IL‐10 productions by isolated PBMC were diminished on ConA‐induced activation but not in response to PHA and anti‐CD3; in CPB patients, only anti‐CD3‐induced IL‐10 production was significantly reduced. Our data indicate that subtle modifications of the reactivity of circulating cells occur during infectious and noninfectious SIRS. Production of both Th1 and Th2 cytokines can be down‐regulated; however, the nature of the SIRS, of the cell population, and of the activator may influence the observation.

CD14/LPS receptor exhibits lectin-like properties

We had previously shown that the specific binding of endotoxin (lipopolysaccharide, LPS) to human monocytes in the presence of serum was mediated by the polysaccharide moiety of the LPS molecule. CD14 has been identified as the main receptor for endotoxin on monocytes/macrophages. In the present report we demonstrate that the CD14 molecule exhibits lectin-like properties. Anti-CD14 monoclonal antibodies inhibited the binding of [3H]-radiolabeled Neisseria meningitidis LPS as efficiently as the homologous unlabeled LPS. Rough Escherichia coli LPS (Rc- and Re-types) could also inhibit the binding of [3H]-LPS to a similar extent, whereas lipid A had no or very weak inhibitory activity. This suggests a major contribution of the inner-core region within the LPS and particularly the Kdo sugars. The lectin-like nature of CD14 was assessed with polyanionic sugars as well as with uncharged polysaccharides. The relative efficiencies in competition were dextran sulfate > fucoidan > mannan > polygalacturonic acid = heparan sulfate ≥ heparin ≥ chondroitin sulfate. Candida albicans phospholipomannan was far more active in the competition experiment than the mannan, indicating that, besides the osidic residues, anionic charges and/or fatty acids may contribute to the interaction with the CD14 molecule. Binding of polysaccharide to CD14 was not sufficient to trigger TNFα and IL-6 production since phospholipomannan and dextran sulfate were unable to induce cytokine release. Taken together, these results demonstrate that the binding of [3H]-LPS to CD14 involves the contribution of sugars and suggest that the signals for cytokine production require additional interactions.

Interleukin-1 receptor antagonist production during infectious and noninfectious systemic inflammatory response syndrome.

Objective To analyze the levels of circulating and cell-associated forms of interleukin-1 receptor antagonist (IL-1ra) and the spontaneous and the lipopolysaccharide- or streptococcus - induced ex vivo production of IL-1ra by isolated neutrophils. Design Cohort study. Setting A collaborative study between an intensive care unit and a research laboratory. Patients Septic patients (those with infectious systemic inflammatory response syndrome [SIRS]) and patients undergoing cardiac surgery with cardiopulmonary bypass (noninfectious SIRS). Measurements and Main Results Both noninfectious and infectious SIRS patients had enhanced levels of plasma IL-1ra. In septic patients, the increased level of IL-1ra associated with circulating leukocytes reflected the higher number of circulating neutrophils, because these cells, as well as peripheral blood mononuclear cells, contained similar levels of cell-associated forms of IL-1ra than those found at homeostasis in healthy controls. The analysis of the in vitro production of IL-1ra by neutrophils showed a decreased capacity of these cells to release the secreted form of IL-1ra on activation in all patients when compared with that capacity in healthy controls. In contrast, the production of the intracellular forms of IL-1ra was not altered in septic patients, but it was diminished in post-cardiopulmonary bypass patients. Conclusions The capacity of releasing IL-1ra by activated neutrophils from infectious or noninfectious SIRS patients was diminished. In contrast, the accumulation of intracellular IL-1ra in septic patients was not modified when compared with that in healthy controls. These ex vivo data illustrate that a different gene regulation of the secreted and intracellular forms of IL-1ra occurs during a pathologic situation like sepsis.

Hyporesponsiveness in leukocytes in sepsis: in vitro models reveal paradoxical effects of IL-10

Sepsis syndrome is linked with a systemic inflammatory response syndrome (SIRS). This severe inflammation is associated with an immune suppression as illustrated by the reduced capacity of circulating leukocytes to produce cytokines in response to in vitro activation. Non-infectious SIRS such as trauma, burn, hemorrhage or major surgery is also associated with a suppression of the immune system. This phenomenon has been recently termed CARS for compensatory anti-inflammatory response syndrome. We report in vitro experiments which suggest that a well-known anti-inflammatory cytokine, namely IL-10, may, in certain experimental conditions, prime the leukocytes finally leading to an increased cytokine production. We discuss the relevance of this in vitro model to the in vivo situations where immune suppression is limited to the blood compartment (or the hematopoietic organs) whereas, in inflammatory foci within the tissues, cytokine production is increased. Our data suggest that IL-10 may be a causative agent of concomitantly occurring SIRS and CARS.