An ex vivo model for studying hepatic schistosomiasis and the effect of released protein from dying eggs

PLoS Negl Trop Dis. 2015 May 12;9(5):e0003760. doi: 10.1371/journal.pntd.0003760. eCollection 2015 May.

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.

Methods and findings: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / immunology*
  • CD40 Antigens / immunology
  • Chemokine CCL4 / immunology
  • Chemokine CXCL5 / immunology
  • Disease Models, Animal
  • Gene Expression
  • Inflammation / immunology
  • Inflammation / parasitology
  • Interleukin-1alpha / immunology
  • Interleukin-1beta / immunology
  • Interleukin-6 / immunology
  • Liver / parasitology*
  • Liver Diseases, Parasitic / immunology*
  • Liver Diseases, Parasitic / parasitology
  • Macrophage Colony-Stimulating Factor / immunology
  • Mice
  • Ovum / immunology*
  • Schistosomiasis / immunology*
  • Schistosomiasis / parasitology
  • Schistosomiasis / pathology
  • Tissue Inhibitor of Metalloproteinase-1 / biosynthesis

Substances

  • Antigens, Protozoan
  • CD40 Antigens
  • Chemokine CCL4
  • Chemokine CXCL5
  • Cxcl5 protein, mouse
  • IL1B protein, mouse
  • Interleukin-1alpha
  • Interleukin-1beta
  • Interleukin-6
  • Tissue Inhibitor of Metalloproteinase-1
  • Macrophage Colony-Stimulating Factor

Grants and funding

The support of the National Health and Medical Research Council of Australia (NHMRC http://www.nhmrc.gov.au) with NHMRC Program Grant (APP1037304) and NHMRC Project Grant (APP1002245) funding is acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.