A Role for TLR4 in Clostridium difficile Infection and the Recognition of Surface Layer Proteins

Clostridium difficile is the etiological agent of antibiotic-associated diarrhoea (AAD) and pseudomembranous colitis in humans. The role of the surface layer proteins (SLPs) in this disease has not yet been fully explored. The aim of this study was to investigate a role for SLPs in the recognition of C. difficile and the subsequent activation of the immune system. Bone marrow derived dendritic cells (DCs) exposed to SLPs were assessed for production of inflammatory cytokines, expression of cell surface markers and their ability to generate T helper (Th) cell responses. DCs isolated from C3H/HeN and C3H/HeJ mice were used in order to examine whether SLPs are recognised by TLR4. The role of TLR4 in infection was examined in TLR4-deficient mice. SLPs induced maturation of DCs characterised by production of IL-12, TNFα and IL-10 and expression of MHC class II, CD40, CD80 and CD86. Furthermore, SLP-activated DCs generated Th cells producing IFNγ and IL-17. SLPs were unable to activate DCs isolated from TLR4-mutant C3H/HeJ mice and failed to induce a subsequent Th cell response. TLR4−/− and Myd88−/−, but not TRIF−/− mice were more susceptible than wild-type mice to C. difficile infection. Furthermore, SLPs activated NFκB, but not IRF3, downstream of TLR4. Our results indicate that SLPs isolated from C. difficile can activate innate and adaptive immunity and that these effects are mediated by TLR4, with TLR4 having a functional role in experimental C. difficile infection. This suggests an important role for SLPs in the recognition of C. difficile by the immune system

CD14 Signaling Restrains Chronic Inflammation through Induction of p38-MAPK/SOCS-Dependent Tolerance

Current thinking emphasizes the primacy of CD14 in facilitating recognition of microbes by certain TLRs to initiate pro-inflammatory signaling events and the importance of p38-MAPK in augmenting such responses. Herein, this paradigm is challenged by demonstrating that recognition of live Borrelia burgdorferi not only triggers an inflammatory response in the absence of CD14, but one that is, in part, a consequence of altered PI3K/AKT/p38-MAPK signaling and impaired negative regulation of TLR2. CD14 deficiency results in increased localization of PI3K to lipid rafts, hyperphosphorylation of AKT, and reduced activation of p38. Such aberrant signaling leads to decreased negative regulation by SOCS1, SOCS3, and CIS, thereby compromising the induction of tolerance in macrophages and engendering more severe and persistent inflammatory responses to B. burgdorferi. Importantly, these altered signaling events and the higher cytokine production observed can be mimicked through shRNA and pharmacological inhibition of p38 activity in CD14-expressing macrophages. Perturbation of this CD14/p38-MAPK-dependent immune regulation may underlie development of infectious chronic inflammatory syndromes

A new process of reconstructing archaeological fires from their impact on sediment: a coupled experimental and numerical approach based on the case study of hearths from the cave of Les Fraux (Dordogne, France)

International audienceA novel approach to the intensity of archaeological fires is proposed, based on a combination of archaeological observations and analyses of sedimentary hearths with relevant proxies obtained from using experimental combustion structures. In this work, two different structures were built and monitored. They aimed at reproducing two types of archaeological hearth morphology encountered at the Bronze Age site of the cave of Les Fraux (Saint Martin de Fressengeas, Dordogne, France). A series of fires was constructed and a large amount of data was collected: temperature curves, wood consumption and observations on substratum evolution. A numerical code for heat transfer was developed to model heat propagation from the surface to the underlying sedimentary layers, the input parameters of which were adapted to fit the thermal evolution observed with the experimental fires. We found that two archaeological parameters are fundamental to characterise the intensity of the fire: the paleotemperature reached at the surface of the burnt sediment (which in our case was determined by thermoluminescence analyses) and the depth of the rubefaction front as an indicator of a 250 °C isothermal surface. We then estimated the duration of an equivalent single fire that would correspond to one of the archaeological hearths investigated. Finally, with the wood consumption recorded during the fire experiments, and the estimated firing duration, the energy involved was evaluated. When generalised to the study of archaeological hearths, this approach could be of great interest in firing intensity evaluation (temperature/time/energy)

Chemical Composition of Polymer Surfaces Imaged by Atomic Force Microscopyand Complementary Approaches

Unfortunately, the editor of this article Hans-Henning Kausch was published as author in the XML-version