Infection of a human hepatoma cell line by hepatitis B virus

Among numerous established human hepatoma cell lines, none has been shown susceptible to hepatitis B virus (HBV) infection. We describe here a cell line, called HepaRG, which exhibits hepatocyte-like morphology, expresses specific hepatocyte functions, and supports HBV infection as well as primary cultures of normal human hepatocytes. Differentiation and infectability are maintained only when these cells are cultured in the presence of corticoids and dimethyl sulfoxide. The specificity of this HBV infection model was ascertained by both the neutralization capacity of HBV-envelope protein-specific antibodies and the competition with an envelope-derived peptide. HepaRG cells therefore represent a tool for deciphering the mechanism of HBV entry. Moreover, their close resemblance to normal human hepatocytes makes them suitable for many applications including drug metabolism studies.

[1]  C. Guguen-Guillouzo,et al.  A plasma membrane protein is involved in cell contact-mediated regulation of tissue-specific genes in adult hepatocytes , 1991, The Journal of cell biology.

[2]  C. Chang,et al.  Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. , 1987, The EMBO journal.

[3]  C. Guguen-Guillouzo,et al.  Role of the Pre-S2 Domain of the Large Envelope Protein in Hepatitis B Virus Assembly and Infectivity , 1998, Journal of Virology.

[4]  M. Frain,et al.  Molecular cloning and sequence analysis of cDNA for human transferrin. , 1984, Biochemical and biophysical research communications.

[5]  C. Ryu,et al.  Generation and characterization of a humanized antibody with specificity for preS2 surface antigen of hepatitis B virus. , 1996, Hybridoma.

[6]  C. Guguen-Guillouzo,et al.  Myristylation of the hepatitis B virus large surface protein is essential for viral infectivity. , 1995, Virology.

[7]  C. Guguen-Guillouzo,et al.  Growth factor dependence of progression through G1 and S phases of adult rat hepatocytes in vitro. Evidence of a mitogen restriction point in mid-late G1. , 1996, The Journal of biological chemistry.

[8]  D. Waxman,et al.  Characterization of rat and human liver microsomal cytochrome P-450 forms involved in nifedipine oxidation, a prototype for genetic polymorphism in oxidative drug metabolism. , 1986, The Journal of biological chemistry.

[9]  M. Frain,et al.  Nucleotide sequence of a cDNA clone for human aldolase B. , 1983, Biochemical and biophysical research communications.

[10]  C. Ryu,et al.  In vitro neutralization of hepatitis B virus by monoclonal antibodies against the viral surface antigen , 1997, Journal of medical virology.

[11]  C. Guguen-Guillouzo,et al.  Infection Process of the Hepatitis B Virus Depends on the Presence of a Defined Sequence in the Pre-S1 Domain , 1999, Journal of Virology.

[12]  S. Law,et al.  Nucleotide sequence and the encoded amino acids of human serum albumin mRNA. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[13]  T. Ochiya,et al.  An in vitro system for infection with hepatitis B virus that uses primary human fetal hepatocytes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[15]  J. Darnell,et al.  Persistence of liver-specific messenger RNA in cultured hepatocytes: different regulatory events for different genes , 1987, The Journal of cell biology.

[16]  C. Guguen-Guillouzo,et al.  A Short N-Proximal Region in the Large Envelope Protein Harbors a Determinant That Contributes to the Species Specificity of Human Hepatitis B Virus , 2001, Journal of Virology.

[17]  C C Howe,et al.  Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. , 1980, Science.

[18]  G. Acs,et al.  Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[19]  C. Guguen-Guillouzo,et al.  Reproducible high level infection of cultured adult human hepatocytes by hepatitis B virus: effect of polyethylene glycol on adsorption and penetration. , 1993, Virology.

[20]  W. Jakoby,et al.  Assays for differentiation of glutathione S-transferases. , 1981, Methods in enzymology.

[21]  C. Bréchot,et al.  Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide , 1988, Journal of virology.

[22]  A. Fautrel,et al.  Viability and function in primary culture of adult hepatocytes from various animal species and human beings after cryopreservation , 1993, Hepatology.

[23]  J. Mullins,et al.  Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA , 1986, Cell.

[24]  S. Urban,et al.  Inhibition of Duck Hepatitis B Virus Infection by a Myristoylated Pre-S Peptide of the Large Viral Surface Protein , 2002, Journal of Virology.

[25]  D. Pompon,et al.  Maximizing the expression of mammalian cytochrome P-450 monooxygenase activities in yeast cells. , 1990, Biochimie.

[26]  B. Geiger,et al.  HBV infection of cell culture: evidence for multivalent and cooperative attachment , 2001, The EMBO journal.

[27]  A. Fujiyama,et al.  Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[28]  K. Koike,et al.  Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[29]  C. Ryu,et al.  Fine mapping of virus-neutralizing epitopes on hepatitis B virus PreS1. , 2000, Virology.

[30]  H. Sambrook.  Molecular cloning : a laboratory manual. Cold Spring Harbor, NY , 1989 .

[31]  H. Zentgraf,et al.  In vitro experimental infection of primary human hepatocytes with hepatitis B virus. , 1994, Gastroenterology.