Thomas Horvatits1, Jan-Erik Wißmann2, Reimar Johne3, Martin H Groschup4, Ashish K Gadicherla3, Julian Schulze Zur Wiesch5, Martin Eiden4, Daniel Todt6, Rudolph Reimer7, Lisa Dähnert4, Anja Schöbel8, Karoline Horvatits9, Rabea Lübke9, Christine Wolschke10, Francis Ayuk10, Meike Rybczynski11, Ansgar W Lohse5, Marylyn M Addo5, Eva Herker8, Marc Lütgehetmann12, Eike Steinmann13, Sven Pischke5. 1. I. Department of Medicine, Gastroenterology and Hepatology, with the Sections Infectious Diseases and Tropical Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems and Heidelberg Partner sites, Germany. Electronic address: t.horvatits@uke.de. 2. Ruhr University Bochum, Faculty of Medicine, Department of Molecular and Medical Virology, Bochum, Germany. 3. German Federal Institute for Risk Assessment, Department Biological Safety, Berlin, Germany. 4. German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems and Heidelberg Partner sites, Germany; Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institute, Greifswald-Isle of Riems, Germany. 5. I. Department of Medicine, Gastroenterology and Hepatology, with the Sections Infectious Diseases and Tropical Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems and Heidelberg Partner sites, Germany. 6. Ruhr University Bochum, Faculty of Medicine, Department of Molecular and Medical Virology, Bochum, Germany; European Virus Bioinformatics Center (EVBC), Jena, Germany. 7. Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany. 8. Institute of Virology, Philipps University Marburg, Marburg, Germany. 9. I. Department of Medicine, Gastroenterology and Hepatology, with the Sections Infectious Diseases and Tropical Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 10. Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 11. University Heart Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 12. German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems and Heidelberg Partner sites, Germany; Institute of Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 13. German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems and Heidelberg Partner sites, Germany; Ruhr University Bochum, Faculty of Medicine, Department of Molecular and Medical Virology, Bochum, Germany.
Abstract
BACKGROUND & AIMS: Hepatitis E virus (HEV) infections are prevalent worldwide. Various viruses have been detected in the ejaculate and can outlast the duration of viremia, indicating replication beyond the blood-testis barrier. HEV replication in diverse organs, however, is still widely misunderstood. We aimed to determine the occurrence, features and morphology of HEV in the ejaculate. METHODS: The presence of HEV in testis was assessed in 12 experimentally HEV-genotype 3-infected pigs. We further tested ejaculate, urine, stool and blood from 3 chronically HEV genotype 3-infected patients and 6 immunocompetent patients with acute HEV infection by HEV-PCR. Morphology and genomic characterization of HEV particles from various human compartments were determined by HEV-PCR, density gradient measurement, immune-electron microscopy and genomic sequencing. RESULTS: In 2 of the 3 chronically HEV-infected patients, we observed HEV-RNA (genotype 3c) in seminal plasma and semen with viral loads >2 logs higher than in the serum. Genomic sequencing showed significant differences between viral strains in the ejaculate compared to stool. Under ribavirin-treatment, HEV shedding in the ejaculate continued for >9 months following the end of viremia. Density gradient measurement and immune-electron microscopy characterized (enveloped) HEV particles in the ejaculate as intact. CONCLUSIONS: The male reproductive system was shown to be a niche of HEV persistence in chronic HEV infection. Surprisingly, sequence analysis revealed distinct genetic HEV variants in the stool and serum, originating from the liver, compared to variants in the ejaculate originating from the male reproductive system. Enveloped HEV particles in the ejaculate did not morphologically differ from serum-derived HEV particles. LAY SUMMARY: Enveloped hepatitis E virus particles could be identified by PCR and electron microscopy in the ejaculate of immunosuppressed chronically infected patients, but not in immunocompetent experimentally infected pigs or in patients with acute self-limiting hepatitis E.
BACKGROUND & AIMS: Hepatitis E virus (HEV) infections are prevalent worldwide. Various viruses have been detected in the ejaculate and can outlast the duration of viremia, indicating replication beyond the blood-testis barrier. HEV replication in diverse organs, however, is still widely misunderstood. We aimed to determine the occurrence, features and morphology of HEV in the ejaculate. METHODS: The presence of HEV in testis was assessed in 12 experimentally HEV-genotype 3-infected pigs. We further tested ejaculate, urine, stool and blood from 3 chronically HEV genotype 3-infected patients and 6 immunocompetent patients with acute HEV infection by HEV-PCR. Morphology and genomic characterization of HEV particles from various human compartments were determined by HEV-PCR, density gradient measurement, immune-electron microscopy and genomic sequencing. RESULTS: In 2 of the 3 chronically HEV-infected patients, we observed HEV-RNA (genotype 3c) in seminal plasma and semen with viral loads >2 logs higher than in the serum. Genomic sequencing showed significant differences between viral strains in the ejaculate compared to stool. Under ribavirin-treatment, HEV shedding in the ejaculate continued for >9 months following the end of viremia. Density gradient measurement and immune-electron microscopy characterized (enveloped) HEV particles in the ejaculate as intact. CONCLUSIONS: The male reproductive system was shown to be a niche of HEV persistence in chronic HEV infection. Surprisingly, sequence analysis revealed distinct genetic HEV variants in the stool and serum, originating from the liver, compared to variants in the ejaculate originating from the male reproductive system. Enveloped HEV particles in the ejaculate did not morphologically differ from serum-derived HEV particles. LAY SUMMARY: Enveloped hepatitis E virus particles could be identified by PCR and electron microscopy in the ejaculate of immunosuppressed chronically infected patients, but not in immunocompetent experimentally infected pigs or in patients with acute self-limiting hepatitis E.
Authors: Johannes H Bannasch; Benjamin Berger; Claus-Peter Schwartkop; Marco Berning; Oliver Goetze; Marcus Panning; Miriam Fritz-Weltin; George Trendelenburg; Mathias Gelderblom; Marc Lütgehetmann; Fridrike Stute; Thomas Horvatits; Meike Dirks; Christoph Antoni; Patrick Behrendt; Sven Pischke Journal: Pathogens Date: 2021-05-30