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IMPACT OF HEV INFECTION ON THE HUMAN HEPATIC INNATE
IMMUNE RESPONSE IN HUMAN LIVER CHIMERIC MICE
1,2
Sayed ,
1
Verhoye ,
Ibrahim M.
Lieven
1
Geert Leroux-Roels ,
3
1
Jacques Izopet , Philip Meuleman
Lander
1
Foquet ,
Florence
3
Abbravanel ,
Ali
1
Farhoudi ,
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.
Transcriptome analysis
Background
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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
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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
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Red: Up regulated gene :
Green: non affected gene
Dotted vs Solid: 4 week , 9 week of HEV gt1
infection respectively
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