Hepatitis B Virus Antigen Development in Cultured Human Hepatocytes

Noyes, Wilbur Fiske

doi:10.3181/00379727-144-37695

Cited by 6 publications

(5 citation statements)

References 12 publications

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“…Although experimental infection of animals with hepadnaviruses can be achieved (3,12,17), one of the major technical problems in hepadnavirus research has been the lack of an in vitro system for productive viral infection and replication. Attempts to infect primary human adult and fetal hepatocyte cultures (2,7,9,21,25), fetal hepatocyte organ cultures (32), and HeLa cells (7) with human HBV have been described. Although intracellular virus-specific proteins were detected after exposure to virus (2,9,23,27) and extracellular virus was found in the culture medium (9), there was no unequivocal evidence for human HBV replication and de novo viral synthesis.…”

mentioning

confidence: 99%

In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus

Tuttleman¹,

Pugh²,

Summers³

1986

J Virol

285

153

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Duck hepatitis B virus (DHBV) obtained from the serum of congenitally infected ducks was used to infect primary duck hepatocyte cultures 1 to 4 days after plating. Virus replication was demonstrated by the appearance, beginning at 2 days after infection, of intracellular covalently closed-circular and single-stranded DHBV DNA replicative intermediates which were not present in the inoculating virus preparation. With increasing time after infection there was further amplification of intracellular relaxed circular, covalently closed-circular, and single-stranded DHBV DNA. Cultures of primary duck hepatocytes are competent for infection with DHBV only during the first 4 days of culture. Synthesis of DHBV core antigen and DHBV surface antigen was detected by imnmunofluorescence in 10% of the hepatocytes in culture. De novo synthesis and release of infectious virus was also demonstrated. Therefore, all stages of viral replication were carried out by these experimentally infected primary hepatocyte cultures. This system makes it possible to study DHBV replication in vitro.

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mentioning

confidence: 99%

In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus

Tuttleman¹,

Pugh²,

Summers³

1986

J Virol

285

153

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“…Propagation of HBV in tissue culture by the use of human embryonic liver cells [Brighton et al, 1971;Zuckerman et al, 1972;Noyes, 1973;Watanabe et al, 19761, organ cultures of liver [Coyne et al, 19711, and human cell line of hepatic origin [Panouse-Perrin et al, 19731 has also been attempted. Though cytoplasmic HBsAg by immunofluorescence or HBsAg in the medium and cell lysates was described in these reports, no clear evidence of HBV replication was demonstrated.…”

Section: Discussionmentioning

confidence: 99%

“…However, further improvement of culture systems is needed for active replication of HBV in vitro. 1973;Panouse-Perrin et al, 1973; Watanabe et al, 19761, mostly by the use of embryonic liver cells, have been unsuccessful so far. Cells used in the previous experiments had no hepatocyte-specific functions such as albumin production.…”

mentioning

confidence: 99%

“…However, all attempts to propagate HBV in cell culture [Brighton et al, 1971; Coyne et al, 1971;Zuckerman et al, 1972;Noyes, and albumin in the cytoplasm of HBV-infected hepatocytes was reported [Michalak et al, 19801. There is, therefore, a possibility that HBV replication is closely related to highly differentiated functions of the host cells.…”

Section: Introductionmentioning

confidence: 99%

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Replication of Hepatitis B Virus in Culture Systems With Adult Human Hepatocytes

Shimizu

Nambu

Kojima

et al. 1986

Journal of Medical Virology

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We developed a technique for isolation and primary culture of adult human hepatocytes from surgical liver biopsy specimens by in situ perfusion and a shaking method. Cultured hepatocytes were maintained in monolayers for more than three weeks and showed morphological and functional characteristics in vivo. The cultured human hepatocytes were inoculated with hepatitis B virus (HBV). Hepatitis B surface antigen (HBsAg) in the medium was detected for about three weeks after inoculation, which was longer than that reported in previous studies. In one case of high attachment efficiency, hepatitis B e antigen (HBeAg) was detected in the medium five to eight days after inoculation. HBsAg and HBeAg were also detected in the extracts of inoculated human hepatocytes. Immunofluorescence study revealed HBsAg in 20-30% of hepatocytes and hepatitis B core antigen (HBcAg) in 2-3% of the cultured human hepatocytes four days after inoculation. Free HBV DNA was identified in the human hepatocytes for at least two weeks after inoculation, although single-stranded HBV DNA was not detected. These studies suggest that HBsAg was actively produced and that HBV replicated in a small number of inoculated adult human hepatocytes in primary culture. However, further improvement of culture systems is needed for active replication of HBV in vitro.

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“…Many important virological experiments with hepatitis B virus (HBV) cannot now be performed because this virus has not been grown in vitro. Although a number of reports of primary isolation have appeared, to date, serial passage of the virus has not been achieved (6,14,20,21).…”

mentioning

confidence: 99%

PLC/PRF/5 (Alexander) hepatoma cell line: further characterization and studies of infectivity

Daemer¹,

Feinstone²,

Alexander³

et al. 1980

Infect Immun

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The Alexander hepatoma cell line, PLC/PRF/5, was studied for evidence of hepatitis B virus markers and alpha-fetoprotein. Only hepatitis B surface antigen and alpha-fetoprotein were detected. Induction experiments with 5-iodo-2'-deoxyuridine and inoculation of chimpanzees with whole cells or tissue culture fluid did not reveal evidence of synthesis of additional hepatitis B virus markers or of production of infectious virus.

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Hepatitis B Virus Antigen Development in Cultured Human Hepatocytes

Cited by 6 publications

References 12 publications

In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus

In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus

Replication of Hepatitis B Virus in Culture Systems With Adult Human Hepatocytes

PLC/PRF/5 (Alexander) hepatoma cell line: further characterization and studies of infectivity

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