Persistent hepatitis C virus infection in microscale primary human hepatocyte cultures

Ploß, Alexander; Khetani, Salman R.; Jones, Christopher T.; Syder, Andrew J.; Trehan, Kartik; Gaysinskaya, Valeriya; Mu, Kathy; Ritola, Kimberly; Rice, Charles M.; Bhatia, Sangeeta N.

doi:10.1073/pnas.0915130107

Cited by 188 publications

(213 citation statements)

References 32 publications

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“…Fetal PHH Long-lived as compared to adult PHH [87][88][89][90]. Structures looking like bile canaliculi [87].…”

Section: Namementioning

confidence: 99%

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Entry and replication of recombinant hepatitis C viruses in cell culture

Vièyres

Pietschmann

2013

Methods

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“…Fetal PHH Long-lived as compared to adult PHH [87][88][89][90]. Structures looking like bile canaliculi [87].…”

Section: Namementioning

confidence: 99%

“…Adult PHH seeded on collagen spots and surrounded by mouse fibroblast "feeder" cells [89,92] Polarized cells, stronger and more stable differentiation than PHH monocultures [92]. [94]…”

Section: Namementioning

confidence: 99%

Entry and replication of recombinant hepatitis C viruses in cell culture

Vièyres

Pietschmann

2013

Methods

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“…contributed equally to this work. falciparum and vivax malaria (13,14). We hypothesized that this system would be ideal for modeling HBV infection in vitro.…”

Section: Significancementioning

confidence: 99%

Modeling host interactions with hepatitis B virus using primary and induced pluripotent stem cell-derived hepatocellular systems

Shlomai

Schwartz

Ramanan³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Hepatitis B virus (HBV) chronically infects 400 million people worldwide and is a leading driver of end-stage liver disease and liver cancer. Research into the biology and treatment of HBV requires an in vitro cell-culture system that supports the infection of human hepatocytes, and accurately recapitulates virus-host interactions. Here, we report that micropatterned cocultures of primary human hepatocytes with stromal cells (MPCCs) reliably support productive HBV infection, and infection can be enhanced by blocking elements of the hepatocyte innate immune response associated with the induction of IFN-stimulated genes. MPCCs maintain prolonged, productive infection and represent a facile platform for studying virus-host interactions and for developing antiviral interventions. Hepatocytes obtained from different human donors vary dramatically in their permissiveness to HBV infection, suggesting that factors-such as divergence in genetic susceptibility to infection-may influence infection in vitro. To establish a complementary, renewable system on an isogenic background in which candidate genetics can be interrogated, we show that inducible pluripotent stem cells differentiated into hepatocyte-like cells (iHeps) support HBV infection that can also be enhanced by blocking interferon-stimulated gene induction. Notably, the emergence of the capacity to support HBV transcriptional activity and initial permissiveness for infection are marked by distinct stages of iHep differentiation, suggesting that infection of iHeps can be used both to study HBV, and conversely to assess the degree of iHep differentiation. Our work demonstrates the utility of these infectious systems for studying HBV biology and the virus' interactions with host hepatocyte genetics and physiology.HBV persistence | innate immunity | viral hepatitis H epatitis B virus (HBV) is a small 3.2-kb DNA virus that selectively infects hepatocytes in the human liver (1). The global disease burden is large, with ∼400 million people chronically infected worldwide, of whom about one-third will develop severe HBV-related complications, such as cirrhosis and liver cancer. Lifelong treatment is often required because of the stable nature of viral episomal DNA, known as covalently closed circular DNA (cccDNA), which maintains basal levels in infected cell nuclei even upon nucleos(t)ide inhibitor treatment. To date, HBV research has been hampered by a distinct lack of robust infectious model systems that both support productive HBV infection and accurately mimic virus-host interactions. Recently, the bile acid pump sodium taurocholate cotransporting polypeptide (NTCP) has been identified as a receptor for both HBV and hepatitis D virus (2), and overexpression of NTCP in hepatoma cell lines renders them susceptible to HBV infection. However, hepatoma cells are known to be defective in many cellular pathways implicated in the innate immune response (3, 4), metabolism (5), and cell proliferation (6), which may contribute to published contradictory evidence regarding the e...

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“…This new experimental model system in which viral titers above 10 5 ffu/ml were achieved allows in addition the validation of antiviral drugs. Improvements of PHH cultivation were also made with the addition of non-parenchymal feeder cells to hepatocytes in micopatterned cocultures (MPCC) (Ploss, Khetani et al 2010). MPCCs displayed hepatic function for several weeks, could be adapted to highthroughput format and were susceptible to HCV infection with lim ited virus spread for cell culture-and patient-derived viruses (Ploss, Khetani et al 2010).…”

Section: Hcv Replication Models In Primary Cells and Patient Isolatesmentioning

confidence: 99%

Cell Culture Systems for Hepatitis C Virus

Steinmann

Pietschmann

2013

Current Topics in Microbiology and Immunology

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Due to the obligatory intracellular life style of viruses, cell culture systems for efficient viral propagation are crucial to obtain a detailed understanding of the virus host cell interaction. For hepatitis C virus (HCV) the development of permissive and authentic culture models has been and continues to be a challenging task. The first ef forts to culture HCV had limited success and range back to before the virus was molecularly cloned in 1989. Since then several major breakthroughs have gradually overcome limitations in culturing the virus and sequentially permitted analysis of viral RNA replication, cell entry and ultimately the complete replication cycle in cultured cells in 2005. Until today, basic and applied HCV research greatly benefit from these tremendous efforts which spurred multiple com plementary cell based model systems for distinct steps of the HCV replication cycle. When used in combination they now permit deep insights into the fascinating biology of HCV and its interplay with the host cell. In fact drug development has been much facilitated and our understanding of the molecular determinants of HCV replication has grown in parallel to these advances. Building on this ground work and further refining our cellular models to better mimic the ar chitecture, polarization and differentiation of natural hepatocytes should reveal novel unique aspects of HCV replication. Ultimately, models to culture primary HCV isolates across all genotypes may teach us important new lessons about viral functional adaptations that have evolved in exchange with its human host and that may ex plain the variable natural course of hepatitis C.

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Persistent hepatitis C virus infection in microscale primary human hepatocyte cultures

Cited by 188 publications

References 32 publications

Entry and replication of recombinant hepatitis C viruses in cell culture

Entry and replication of recombinant hepatitis C viruses in cell culture

Modeling host interactions with hepatitis B virus using primary and induced pluripotent stem cell-derived hepatocellular systems

Cell Culture Systems for Hepatitis C Virus

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