Factors determining successful engraftment of hepatocytes and susceptibility to hepatitis B and C virus infection in uPA-SCID mice

Vanwolleghem, Thomas; Libbrecht, Louis; Hansen, Bettina E.; Desombere, Isabelle; Roskams, Tania; Meuleman, Philip; Leroux‐Roels, Geert

doi:10.1016/j.jhep.2010.03.024

Cited by 68 publications

(86 citation statements)

References 26 publications

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“…Interestingly, after inoculation with HCV-positive sera of different genotypes, none of the mice exhibited a detectable level of HCV RNA in the serum at 2 weeks pi, and only about half of them exhibited a detectable level at 1 month pi. This is in contrast with what has been described after inoculation of chimeric mice engrafted with PHHs (25,26,58,59), where all the engrafted mice inoculated with HCV exhibit detectable viral titers at early time points. Our data show that only mice with hALB titers over 450 μg/ml at the time of HCV inoculation, equivalent to a liver repopulation around 5%, had detectable HCV RNA 1 month pi.…”

Section: Methodscontrasting

confidence: 99%

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

et al. 2014

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Section: Methodscontrasting

confidence: 99%

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

et al. 2014

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“…2A), which correlated with liver repopulation by human hepatocytes as demonstrated by H&E, specific human albumin, and human mitochondrial staining (Fig. 2B) (12). To corroborate the observed feces-derived HEV in vitro infectivity, filtered undiluted fecal suspensions (see Materials and Methods) from patients HEV0069 and HEV0122 were i.v.…”

Section: Resultssupporting

confidence: 58%

“…uPA homozygous mice, 6 to 12 weeks of age, were transplanted as previously described (12). In short, mice were anesthetized and transplanted via intrasplenic injection with 0.5 ϫ 10 6 to 2 ϫ 10 6 viable commercially available cryopreserved human hepatocytes (Lonza, Basel, Switzerland, and Corning, Corning, NY, USA).…”

Section: Methodsmentioning

confidence: 99%

“…In short, mice were anesthetized and transplanted via intrasplenic injection with 0.5 ϫ 10 6 to 2 ϫ 10 6 viable commercially available cryopreserved human hepatocytes (Lonza, Basel, Switzerland, and Corning, Corning, NY, USA). Graft take was determined by human albumin in mouse serum using an ELISA with human albumin cross-adsorbed antibody (Bethyl laboratories, Montgomery, TX, USA) as previously described (12).…”

Section: Methodsmentioning

confidence: 99%

“…Human liver chimeric mice have contributed significantly to our understanding of viral pathobiology, virus-host interactions, and antiviral therapy for hepatitis B, C, and D infections (12)(13)(14)(15)(16)(17)(18). Since no animal model for chronic HEV infection is available, we examined the infectivity of HEV gt3 samples of different clinical origins in the humanized liver urokinase-type plasminogen activator (uPA) transgenic mouse model on a severely immunodeficient NOD/Shi-scid/IL-2R␥ null background (uPA-NOG) and compared this to the established adenocarcinoma human alveolar basal epithelial cells (A549) culture system (19)(20)(21)(22).…”

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Hepatitis E Virus (HEV) Genotype 3 Infection of Human Liver Chimeric Mice as a Model for Chronic HEV Infection

Garde

Pas

Net

et al. 2016

J Virol

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Genotype 3 (gt3) hepatitis E virus (HEV) infections are emerging in Western countries. Immunosuppressed patients are at risk of chronic HEV infection and progressive liver damage, but no adequate model system currently mimics this disease course. Here we explore the possibilities of in vivo HEV studies in a human liver chimeric mouse model (uPA ؉/؉ Nod-SCID-IL2R␥ ؊/؊ ) next to the A549 cell culture system, using HEV RNA-positive EDTA-plasma, feces, or liver biopsy specimens from 8 immunocompromised patients with chronic gt3 HEV. HEV from feces-or liver-derived inocula showed clear virus propagation within 2 weeks after inoculation onto A549 cells, compared to slow or no HEV propagation of HEV RNA-positive, EDTA-plasma samples. These in vitro HEV infectivity differences were mirrored in human-liver chimeric mice after intravenous (i.v.) inoculation of selected samples. HEV RNA levels of up to 8 log IU HEV RNA/gram were consistently present in 100% of chimeric mouse livers from week 2 to week 14 after inoculation with human feces-or liver-derived HEV. Feces and bile of infected mice contained moderate to large amounts of HEV RNA, while HEV viremia was low and inconsistently detected. Mouse-passaged HEV could subsequently be propagated for up to 100 days in vitro. In contrast, cell culture-derived or seronegative EDTA-plasma-derived HEV was not infectious in inoculated animals. In conclusion, the infectivity of feces-derived human HEV is higher than that of EDTA-plasma-derived HEV both in vitro and in vivo. Persistent HEV gt3 infections in chimeric mice show preferential viral shedding toward mouse bile and feces, paralleling the course of infection in humans. H epatitis E virus (HEV) is a nonenveloped positive-sense single-stranded RNA virus of the genus Orthohepevirus and family Hepeviridae (1). Four major HEV genotypes infecting humans have been described so far. Genotype 1 (gt1) and gt2 strains are isolated only from humans, whereas gt3 and gt4 strains are considered zoonotic viruses, present in both humans and several other species like pigs and wild game. HEV is spread through the oral-fecal route via contaminated water in developing countries or, among other routes, via direct contact with animals or the consumption of undercooked meat in industrialized countries. In immunocompetent individuals, HEV infection is mainly self-limiting and often asymptomatic and thus remains largely underdiagnosed. HEV infections in immunosuppressed patients, such as solid-organ transplant recipients, often persist and can progress quickly to liver fibrosis and cirrhosis (2-4).HEV gt3 infections are emerging in Western countries, including France, the United Kingdom, and the Netherlands (5-7). Despite an overall decrease in anti-HEV seroprevalence from 1996 to 2011, young adult blood donors demonstrated higher seroprevalences from 2000 to 2011 (8, 9). In addition, HEV RNA-positive blood donations were reported to increase in the Netherlands since 2012 (10). Although the exact source of this HEV gt3 infection is unknown,...

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Responsive polymer‐assisted 3D cryogel supports Huh7.5 as in vitro hepatitis C virus model and ectopic human hepatic tissue in athymic mice

Jayal

Behera

Mullick

et al. 2020

Biotech & Bioengineering

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The three‐dimensional (3D) cell culture models serve as the interface between conventional two‐dimensional (2D) monolayer culture and animal models. 3D culture offers the best possible model system to understand the pathophysiology of human pathogens such as hepatitis C virus (HCV), which lacks a small animal model, due to narrow host tropism and non‐permissiveness of murine hepatocytes. In this study, functionally robust spheroids of HCV permissive Huh7.5 cells were generated, assisted by the temperature or pH‐responsive polymers PNIPAAm and Eudragit respectively, followed by the long‐term growth of the multilayered 3D aggregates in poly(ethylene glycol) (PEG)–alginate–gelatin (PAG) cryogel. The human serum albumin (HSA), marker of hepatic viability was detected up to 600 ng/ml on 24th day of culture. The 3D spheroid culture exhibited a distinct morphology and transcript levels with the upregulation of hepato‐specific transcripts, nuclear factor 4α (HNF4α), transthyretin (TTr), albumin (Alb), phase I and phase II drug‐metabolizing genes. The two most important phase I enzymes CYP3A4 and CYP2D6, together responsible for 90% metabolism of drugs exhibited up to 9‐ and 12‐fold increment, respectively in transcripts. The 3D culture was highly permissive to HCV infection and supported higher multiplicity of infection compared to monolayer Huh7.5 culture. Quantitation of high levels of HSA (500–200 ng/ml) in circulation in mice for 32 days asserted integration with host vasculature and in vivo establishment of 3D culture implants as an ectopic human hepatic tissue in mice. The study demonstrates the 3D spheroid Huh7.5 culture as a model for HCV studies and screening potential for anti‐HCV drug candidates.

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Factors determining successful engraftment of hepatocytes and susceptibility to hepatitis B and C virus infection in uPA-SCID mice

Cited by 68 publications

References 26 publications

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

Hepatitis E Virus (HEV) Genotype 3 Infection of Human Liver Chimeric Mice as a Model for Chronic HEV Infection

Responsive polymer‐assisted 3D cryogel supports Huh7.5 as in vitro hepatitis C virus model and ectopic human hepatic tissue in athymic mice

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