Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection
Xiping Zhao, … , Hubert E. Blum, Thomas F. Baumert
Xiping Zhao, … , Hubert E. Blum, Thomas F. Baumert
Published January 15, 2002
Citation Information: J Clin Invest. 2002;109(2):221-232. https://doi.org/10.1172/JCI13011.
View: Text | PDF
Article

Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection

Abstract

Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide, but the study of HCV infection has been hampered by the lack of an in vitro or in vivo small animal model. The tree shrew Tupaia belangeri is susceptible to infection with a variety of human viruses in vivo, including hepatitis viruses. We show that primary Tupaia hepatocytes can be infected with serum- or plasma-derived HCV from infected humans, as measured by de novo synthesis of HCV RNA, analysis of viral quasispecies evolution, and detection of viral proteins. Production of infectious virus could be demonstrated by passage to naive hepatocytes. To assess whether viral entry in Tupaia hepatocytes was dependent on the recently isolated HCV E2 binding protein CD81, we identified and characterized Tupaia CD81. Sequence analysis of cloned Tupaia cDNA revealed a high degree of homology between Tupaia and human CD81 large extracellular loops (LEL). Cellular binding of E2 and HCV infection could not be inhibited by anti-CD81 antibodies or soluble CD81-LEL, suggesting that viral entry can occur through receptors other than CD81. Thus, primary Tupaia hepatocytes provide a potential model for the study of HCV infection of hepatocytes.

Authors

Xiping Zhao, Zhen-Ya Tang, Bettina Klumpp, Guido Wolff-Vorbeck, Heidi Barth, Shoshana Levy, Fritz von Weizsäcker, Hubert E. Blum, Thomas F. Baumert

×

Figure 2

Options: View larger image (or click on image) Download as PowerPoint
Passage of HCV in primary Tupaia hepatocytes. (a) Passage of HCV to naiv...
Passage of HCV in primary Tupaia hepatocytes. (a) Passage of HCV to naive cells. Primary Tupaia hepatocytes were incubated with HCV RNA–positive plasma or serum (1: 10 μl H77 7-12-77, 10–1 dilution; 2: 10 μl ES-2; or 3: 50 μl A5387) 1 day after plating. 5 days later, HCV infection was determined by RT-PCR of cellular positive- and negative-strand HCV RNA, on day 5 after incubation with serum-derived HCV, as described above. Strand-specificity of RT-PCR was assessed using 10 fg of in vitro–synthesized positive- (lanes 1 and 7) and negative-strand HCV RNA (lanes 2 and 8). On day 5 after infection, medium was transferred to naive hepatocytes, and infection was assessed 5 days later as described above. (b) Serial passage. Primary Tupaia hepatocytes were infected with serum 3 as described above. On day 5 after infection, medium was transferred to naive hepatocytes from a separate preparation (first passage). Eight days later, medium was used to infect a third preparation of hepatocytes (second passage). Infection of hepatocytes was assessed as described above.