title: An ex vivo model for studying hepatic schistosomiasis and the effect of released protein from dying eggs.
abstract: BACKGROUND
  We report the use of an ex vivo precision cut liver slice (PCLS) mouse model for studying hepatic schistosomiasis. In this system, liver tissue is unfixed, unfrozen, and alive for maintenance in culture and subsequent molecular analysis.
  
  
  RESULTS
  Using thick naive mouse liver tissue and sterile culture conditions, the addition of soluble egg antigen (SEA) derived from Schistosoma japonicum eggs, followed 4, 24 and 48 hrs time points. Tissue was collected for transcriptional analysis and supernatants collected to quantitate liver enzymes, cytokines and chemokines. No significant hepatotoxicity was demonstrated by supernatant liver enzymes due to the presence of SEA. A proinflammatory response was observed both at the transcriptional level and at the protein level by cytokine and chemokine bead assay. Key genes observed elevated transcription in response to the addition of SEA included: IL1-α and IL1-β, IL6, all associated with inflammation. The recruitment of antigen presenting cells was reflected in increases in transcription of CD40, CCL4 and CSF1. Indications of tissue remodeling were seen in elevated gene expression of various Matrix MetalloProteinases (MMP3, 9, 10, 13) and delayed increases in TIMP1. Collagen deposition was significantly reduced in the presence of SEA as shown in COL1A1 expression by qPCR after 24 hrs culture. Cytokine and chemokine analysis of the culture supernatants confirmed the elevation of proteins including IL6, CCL3, CCL4 and CXCL5.
  
  
  CONCLUSIONS
  This ex vivo model system for the synchronised delivery of parasite antigen to liver tissue provides an insight into the early phase of hepatic schistosomiasis, corresponding with the release of soluble proteins from dying schistosome eggs.
authors.affiliation: QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Geoffrey N
authors.initials: GN
authors.lastname: Gobert
authors.affiliation: QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Sujeevi K
authors.initials: SK
authors.lastname: Nawaratna
authors.affiliation: QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Marina
authors.initials: M
authors.lastname: Harvie
authors.affiliation: QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Grant A
authors.initials: GA
authors.lastname: Ramm
authors.affiliation: QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Donald P
authors.initials: DP
authors.lastname: McManus
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology; D054407:Chemokine CCL4 / Q000276:immunology; D054365:Chemokine CXCL5 / Q000276:immunology; D004195:Disease Models, Animal; D015870:Gene Expression; D007249:Inflammation / Q000276:immunology / Q000469:parasitology; D053582:Interleukin-1alpha / Q000276:immunology; D053583:Interleukin-1beta / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D008099:Liver / Q000469:parasitology*; D008109:Liver Diseases, Parasitic / Q000276:immunology* / Q000469:parasitology; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology; D051379:Mice; D010063:Ovum / Q000276:immunology*; D012552:Schistosomiasis / Q000276:immunology* / Q000469:parasitology / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis
chemical_list: D000953:Antigens, Protozoan; D019013:CD40 Antigens; D054407:Chemokine CCL4; D054365:Chemokine CXCL5; C094739:Cxcl5 protein, mouse; C587672:IL1B protein, mouse; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D015850:Interleukin-6; D019715:Tissue Inhibitor of Metalloproteinase-1; D016173:Macrophage Colony-Stimulating Factor