Visualization of hepatitis E virus RNA and proteins in the human liver

JOURNAL OF HEPATOLOGY

卷:67

Article

Lenggenhager, Daniela^1,2  Gouttenoire, Jerome³  Malehmir, Mohsen^1,2  Bawohl, Marion^1,2  Honcharova-Biletska, Hanna^1,2  Kreutzer, Susanne^1,2  Semela, David⁴  Neuweiler, Jorg⁵  Hurlimann, Sandra⁶  Aepli, Patrick⁷  Fraga, Montserrat³  Sahli, Roland⁸  Terracciano, Luigi⁹  Rubbia-Brandt, Laura¹⁰  Mullhaupt, Beat^11,12  Sempoux, Christine¹³  Moradpour, Darius³  Weber, Achim^1,2 

[1] Univ Zurich, Dept Pathol & Mol Pathol, Schmelzbergstr 12, CH-8091 Zurich, Switzerland

[2] Univ Hosp Zurich, Schmelzbergstr 12, CH-8091 Zurich, Switzerland

[3] Univ Lausanne, Div Gastroenterol & Hepatol, CHU Vaudois, Lausanne, Switzerland

[4] Cantonal Hosp St Gallen, Div Gastroenterol & Hepatol, St Gallen, Switzerland

[5] Cantonal Hosp St Gallen, Inst Pathol, St Gallen, Switzerland

[6] Cantonal Hosp Lucerne, Inst Pathol, Luzern, Switzerland

[7] Cantonal Hosp Lucerne, Gastroenterol & Hepatol Unit, Luzern, Switzerland

[8] Univ Lausanne, Inst Microbiol, CHU Vaudois, Lausanne, Switzerland

[9] Univ Hosp Basel, Dept Pathol, Basel, Switzerland

[10] Geneva Univ Hosp, Fac Med, Serv Pathol Clin, Geneva, Switzerland

[11] Univ Zurich, Clin Hepatol, Zurich, Switzerland

[12] Univ Zurich, Clin Gastroenterol, Zurich, Switzerland

[13] Univ Lausanne, Inst Univ Pathol, CHU Vaudois, Lausanne, Switzerland

关键词: Hepatitis E virus (HEV);    Histopathology;    Immunohistochemistry;    In situ hybridization;    Hepatitis;    Human liver;   

DOI  :  10.1016/j.jhep.2017.04.002

来源: Elsevier

【 摘 要 】

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. (C) 2017 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

【 授权许可】

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