1. Impact of HEV infection on the human hepatic innate immune response in human liver chimeric mice
Ibrahim M. Sayed1,2, Lieven Verhoye1, Lander Foquet1, Florence Abbravanel3, Ali Farhoudi1, Geert Leroux-Roels 1,
Jacques Izopet3, Philip Meuleman1
1Laboratory of Liver Infectious Diseases, Department of Clinical Chemistry, Microbiology and Immunology, UGENT, Ghent, Belgium.
2Microbiology and Immunology Department, Assiut University, Assiut, Egypt.
3INSERM U1043, IFR-BMT, CHU Purpan, Toulouse, France; Université Paul-Sabatier, Toulouse, France; Laboratory of Virology, CHU Purpan, Toulouse, France.
Background
Transcriptome analysis
Every year approximately 20 million people become infected with HEV
HEV causes acute, chronic and extra-hepatic manifestation
Transmission routes: fecal-oral, foodborne, vertical, blood transfusion
Lack of robust cell culture systems covering all HEV genotypes
Non human primates are susceptible to HEV gt1-4, Pigs are susceptible to HEV gt3-4
Human liver chimeric uPA/SCID mice are susceptible to HEV infection
Aim: Study the impact of HEV infection on hepatic innate immune response of human liver chimeric mice
Experimental protocol
Mice
Humanized uPA+/+-SCID mice infected with HEV
Non infected humanized uPA+/+-SCID mice
HEV inocula:
Stool suspension of infected chimpanzee (genotype 1, Sar-55 strain)
Stool from infected patient (genotype 3f)
Analysis
Detect and quantify HEV RNA in mouse plasma and stool
Liver analysis: quantification HEV RNA inside mouse liver
Gene expression
HEV gt1 and HEV gt3 infected liver
Liver fragments from freshly sacrificed mice
RT-qPCR & human-specific primer/probe combinations
Non-inoculated humanized mice as negative control
Five house keeping genes (stability using GeNom)
Up-/down-regulated gene: fold change ≥2
Infection of humanized mice with HEV
HEV gt1 is more virulent than HEV gt3
Transcriptome analysis
Conclusion
Human liver chimeric mice can be used to study HEV pathogenesis
Numerous genes are upregulated upon HEV gt1 infection
Up-regulation was more pronounced at later time points during infection
Only minimal upregulation upon HEV gt3 infection
Further research is needed to define the factors responsible for different profile between gt1 and gt3
Red: Up regulated gene :
Green: non affected gene
Dotted vs Solid: 4 week , 9 week of HEV gt1 infection respectively