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Ultrastructural organisation of HCV from the bloodstream of infected patients revealed by electron microscopy after specific immunocapture
  1. Eric Piver1,2,
  2. Audrey Boyer1,3,
  3. Julien Gaillard4,
  4. Anne Bull1,
  5. Elodie Beaumont1,
  6. Philippe Roingeard1,4,
  7. Jean-Christophe Meunier1
  1. 1INSERM U966, Faculté de Médecine, Université François Rabelais and CHRU de Tours, Tours, France
  2. 2Biochimie & Biologie Moléculaire, Hôpital Trousseau, CHRU de Tours, Tours, France
  3. 3Liang Laboratory, Liver Diseases Branch, NIDDK, National Institutes of Health, Bethesda, Maryland, USA
  4. 4Plate-Forme IBiSA des Microscopies, PPF ASB, Université François Rabelais and CHRU de Tours, Tours, France
  1. Correspondence to Dr Jean-Christophe Meunier, INSERM U966, Faculté de Médecine, Université François Rabelais, 10 Bd Tonnellé—BP 3223, Tours Cedex 37032, France; jean.meunier{at}univ-tours.fr

Abstract

Objective HCV particles are associated with very low-density lipoprotein components in chronically infected patients. These hybrid particles, or ‘lipo-viro particles’ (LVPs), are rich in triglycerides, and contain the viral RNA, the capsid protein, E1E2 envelope glycoproteins and apolipoproteins B and E. However, their specific ultrastructural organisation has yet to be determined. We developed a strategy for the preparation of any viral sample that preserves the native structure of the LVPs, facilitating their precise morphological characterisation.

Design Using a strategy based on the direct specific immunocapture of particles on transmission electron microscopy (TEM) grids, we characterised the precise morphology of the viral particle by TEM.

Results The LVP consists of a broad nucleocapsid surrounding an electron-dense centre, presumably containing the HCV genome. The nucleocapsid is surrounded by an irregular, detergent-sensitive crescent probably composed of lipids. Lipid content may determine particle size. These particles carry HCV E1E2, ApoB and ApoE, as shown in our immuno-EM analysis. Our results also suggest that these putative LVPs circulate in the serum of patients as part of a mixed population, including lipoprotein-like particles and complete viral particles.

Conclusions Twenty-five years after the discovery of HCV, this study finally provides information about the precise morphological organisation of viral particles. It is truly remarkable that our TEM images fully confirm the ultrastructure of LVPs predicted by several authors, almost exclusively from the results of molecular biology studies.

  • LIPID METABOLISM
  • INFECTIOUS DISEASE
  • IMAGING
  • HEPATITIS C
  • HCV

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Footnotes

  • AB and JG contributed equally.

  • Contributors EP and J-CM planned and performed experiments, and wrote the manuscript. JG, ABo, ABu performed experimental work; EB provided reagents. PR provided the laboratory infrastructure and edited the manuscript.

  • Funding This work was supported by an ‘Initiatives Académiques’ grant from Région Centre, France. This work was also supported by a 12–24-month research grant from the ANRS (Agence Nationale pour la Recherche sur le Sida et les hépatites virales), France.

  • Competing interests None declared.

  • Patient consent Obtained.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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