A mouse model of post-stroke pneumonia induced by intra-tracheal inoculation with Streptococcus pneumoniae

Background: Stroke-induced immunodeficiency increases the risk of infectious complications, which adversely affects neurological outcome. Among those, pneumonia affects as many as one third of stroke patients and is the main contributor to mortality in the post-acute phase of stroke. Experimental findings on post-stroke susceptibility to spontaneous pneumonia in mice are contradictory. Here, we established a mouse model inducing standardized bacterial pneumonia and characterized the impaired pulmonary cellular and humoral immune responses after experimental stroke. Methods: Bacterial pneumonia was induced by intra-tracheal inoculation with Streptococcus pneumoniae at different time points after transient middle cerebral artery occlusion (MCAO). Bacterial counts in lungs and blood, histological changes, and cytokine production in the lungs were assessed. Furthermore, we investigated the effect of pneumonia on stroke outcome. Results: Intra-tracheal inoculation resulted in reproducible pneumonia and bacteraemia, and demonstrated post-stroke susceptibility to streptococcal pneumonia developing with a delay of at least 24 h after MCAO. Higher bacterial counts in mice infected 3 days after stroke induction correlated with reduced neutrophil and macrophage infiltration in the lungs and lower levels of pro-inflammatory cytokines in the broncho-alveolar lavage compared to sham-operated animals. Pneumonia increased mortality without affecting brain-infiltrating leukocytes. Conclusions: In this standardized mouse model of post-stroke pneumonia, we describe attenuated leukocyte infiltration and cytokine production in response to bacterial infection in the lungs that has a profound effect on outcome

A complementary study approach unravels novel players in the pathoetiology of Hirschsprung disease

Hirschsprung disease (HSCR, OMIM 142623) involves congenital intestinal obstruction caused by dysfunction of neural crest cells and their progeny during enteric nervous system (ENS) development. HSCR is a multifactorial disorder; pathogenetic variants accounting for disease phenotype are identified only in a minority of cases, and the identification of novel disease-relevant genes remains challenging. In order to identify and to validate a potential disease-causing relevance of novel HSCR candidate genes, we established a complementary study approach, combining whole exome sequencing (WES) with transcriptome analysis of murine embryonic ENS-related tissues, literature and databas

Osteoclast Generation and Cytokine Profile at Prosthetic Interfaces: A Study on Tissue of Patients with Aseptic Loosening or Implant-Associated Infections

Aseptic loosening of implants or loosening due to persistent bacterial infection remains a severe complication in orthopaedic surgery. To investigate underlying cellular and molecular mechanisms, particularly with regard to bone loss, tissue samples of patients requiring surgery were examined. By histological methods and by quantitative RT-PCR, respectively, infiltration of leukocytes, expression of osteoclast-typical genes and of proinflammatory cytokines was determined. Samples were taken directly from osteolytic sites and for comparison from adjacent sites, distant sites and from muscle. At osteolytic sites, cathepsin K and the metalloproteinases MMP1 and MMP9 were found, as was expression of inflammation-related cytokines, particularly of interleukin (IL)-1β, CXCL8, S100A9 and a very moderate expression of receptor activator of NfκB ligand (RANKL) and tumour necrosis factor (TNF) a. Of note, expression of these parameters gradually decreased from sites of osteolysis to adjacent tissue, to distant tissue to muscle. In patients with infection and osteolysis, expression of cytokines, notably of CXCL8, was markedly enhanced, especially in adjacent and distant tissues, where expression was 10- to 20-fold higher compared to tissue of aseptic patients. A possible source of CXCL8 could be infiltrated cells, particularly neutrophils, because they were found in infected tissue only. Histological examination of the biopsies revealed an additional CXCL8 source, namely endothelial cells of small blood vessels. In conclusion, aseptic loosening and implant-associated infection are associated with osteoclast generation and a local inflammatory response. The proinflammatory environment could promote the differentiation of precursor cells to osteoclasts, thereby linking inflammation to bone resorption. The higher expression of cytokines, particularly of CXCL8 in tissue of patients with bacterial infection, could explain the accelerated time course of bone resorption as it occurs in infection compared to aseptic loosening

Functional characterisation of decoy receptor 3 in Crohn's disease

AIMS: Both epithelial barrier dysfunction and apoptosis resistance of immune cells contribute to the pathogenesis of Crohn's disease. The soluble decoy receptor 3 (DcR3) acts in an anti-apoptotic manner by neutralising the death ligand CD95L. Here, we investigated the possible involvement of DcR3 in Crohn's disease. METHODS: The epithelial fraction of human small intestinal mucosa samples was obtained by laser microdissection. Expression of DcR3 was examined by global gene expression profiling, quantitative reverse transcription polymerase chain reaction, immunoblot analysis, and immunohistochemistry. DcR3 concentrations in the serum of patients with Crohn's disease were measured by enzyme-linked immunosorbent assay. Apoptosis assays were performed to study the effects of DcR3 in intestinal epithelial cells and lamina propria T cells. RESULTS: DcR3 is over-expressed in the epithelial layer of ileum specimens in patients with Crohn's disease, both at actively inflamed and non-active sites. DcR3 serum levels are significantly elevated in patients with active and non-active Crohn's disease as compared to healthy controls. The expression of DcR3 in intestinal epithelial cells is induced by tumour necrosis factor alpha. Increased DcR3 expression is associated with activation of nuclear factor kappa B (NF-kappaB) and results in protection of intestinal epithelial cells and lamina propria T cells from CD95L-induced apoptosis. CONCLUSIONS: DcR3 may promote inflammation in Crohn's disease by inhibiting CD95L-induced apoptosis of epithelial and immune cells as well as by inducing NF-kappaB activation