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Visualization of hepatitis E virus RNA and proteins in the human liver
- Source :
- Journal of Hepatology, Vol. 67, No 3 (2017) pp. 471-479
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- Background & Aims Although hepatitis E constitutes a substantial disease burden worldwide, surprisingly little is known about the localization of hepatitis E virus (HEV) in the human liver. We therefore aimed to visualize HEV RNA and proteins in situ . Methods A panel of 12 different antibodies against HEV open reading frame (ORF) 1–3 proteins was evaluated for immunohistochemistry (IHC) and two probes for in situ hybridization (ISH) in formalin-fixed, paraffin-embedded (FFPE) HuH7 cells transfected with HEV ORF1-3 expression vectors. IHC (and partly ISH) were then applied to Hep293TT cells replicating infectious HEV and liver specimens from patients with hepatitis E (n=20) and controls (n=134). Results Whereas ORF1-3 proteins were all detectable in transfected, HEV protein-expressing cells, only ORF2 and 3 proteins were traceable in cells replicating infectious HEV. Only the ORF2-encoded capsid protein was also unequivocally detectable in liver specimens from patients with hepatitis E. IHC for ORF2 protein revealed a patchy expression in individual or grouped hepatocytes, generally stronger in chronic compared to acute hepatitis. Besides cytoplasmic and canalicular, ORF2 protein also displayed a hitherto unknown nuclear localization. Positivity for ORF2 protein in defined areas correlated with HEV RNA detection by ISH. IHC was specific and comparably sensitive as PCR for HEV RNA. Conclusions ORF2 protein can be reliably visualized in the liver of patients with hepatitis E, allowing for sensitive and specific detection of HEV in FFPE samples. Its variable subcellular distribution in individual hepatocytes of the same liver suggests a redistribution of ORF2 protein during infection and interaction with nuclear components. Lay summary The open reading frame (ORF) 2 protein can be used to visualize the hepatitis E virus (HEV) in the human liver. This enabled us to discover a hitherto unknown localization of the HEV ORF2 protein in the nucleus of hepatocytes and to develop a test for rapid histopathologic diagnosis of hepatitis E, the most common cause of acute hepatitis worldwide.
- Subjects :
- 0301 basic medicine
viruses
610 Medicine & health
In situ hybridization
ddc:616.07
medicine.disease_cause
Viral Proteins
03 medical and health sciences
Hepatitis E virus
Cell Line, Tumor
10049 Institute of Pathology and Molecular Pathology
medicine
Humans
In Situ Hybridization
Hepatitis
Expression vector
Hepatology
biology
virus diseases
RNA
Hepatitis E
medicine.disease
Immunohistochemistry
Virology
digestive system diseases
10219 Clinic for Gastroenterology and Hepatology
030104 developmental biology
Liver
Capsid
Tissue Array Analysis
biology.protein
RNA, Viral
2721 Hepatology
Antibody
Subjects
Details
- ISSN :
- 01688278
- Volume :
- 67
- Database :
- OpenAIRE
- Journal :
- Journal of Hepatology
- Accession number :
- edsair.doi.dedup.....d7f5d31b4bab5b620cd6a2c9da0c7086