Sustained survival of human hepatocytes in mice: A model for in vivo infection with human hepatitis B and hepatitis delta viruses

Abstract

Persistence of hepatocytes transplanted into the same or related species has been established. The long-term engraftment of human hepatocytes into rodents would be useful for the study of human viral hepatitis, where it might allow the species, technical and size limitations of the current animal models to be overcome. Although transgenic mice expressing the hepatitis B virus (HBV) genome produce infectious virus in their serum, the viral life cycle is not complete, in that the early stages of viral binding and entry into hepatocytes and production of an episomal transcriptional DNA template do not occur. As for hepatitis delta virus (HDV), another cause of liver disease, no effective therapy exists to eradicate infection, and it remains resistant even to recent regimens that have considerably changed the treatment of HBV (ref. 13). Here, we demonstrate long-term engraftment of primary human hepatocytes transplanted in a matrix under the kidney capsule of mice with administration of an agonistic antibody against c-Met. These mice were susceptible to HBV infection and completion of the viral life cycle. In addition, we demonstrate super-infection of the HBV-infected mice with HDV. Our results describe a new xenotransplant model that allows study of multiple aspects of human hepatitis viral infections, and may enhance studies of human liver diseases.

Heterotopically Transplanted Hepatocyte Survival Depends on Extracellular Matrix Components

“…1,3 In the liver, hepatocytes are surrounded by ECMs that are important for functional and structural maintenance through cell-cell and cell-ECM interactions. 8 However, once hepatocytes are isolated from their natural environment, they detach from the ECMs and undergo apoptosis through detachment-induced cell death that occurs within a short period after isolation.…”

“…Since the marker protein of the transplanted hepatocytes, hAAT, demonstrates a short half-life (less than 2 hours) in the mouse 7 and is produced only from the hepatocytes, the viability and survival of the transplanted hepatocytes in vivo can reasonably be assessed by periodic mouse serum measurement of the hAAT reporter protein, as described previously. [3][4][5][6][7] Mice were sacrificed at day 140; the grafts were excised and processed for histological examination.…”

“…1,2 We recently reported that the inefficient hepatocyte survival may be overcome by providing ECMs extracted from murine Engelbreth-Holm-Swarm tumor cell lines (EHS-ECMs). [3][4][5][6] The EHS-ECMs are rich in ECM components, predominantly laminin and type IV collagen, but also contain a small amount of growth factors, such as epidermal growth factor. This study was performed to determine the effects of EHS-ECMs on hepatocyte survival and whether it was solely due to ECMs or to a combination of ECMs and growth factors in EHS-ECMs.…”

Repopulation of Mouse Liver With Human Hepatocytes And In Vivo Infection With Hepatitis B Virus

“…Clearly, our system will offer the possibility to test antivirals that interfere not only with viral replication, but also with viral spread. In 2 other recently reported xenotransplantation models, cell engraftment was achieved by implanting human liver slices 42 or hepatocyte suspensions 43 extrahepatically, under the kidney capsule of immunodeficient mice. Although production of HBV was demonstrated, both these systems allowed survival, but not growth, of human hepatocytes in mice.…”

Improved gene transfer selectivity to hepatocarcinoma cells by retrovirus vector displaying single-chain variable fragment antibody against c-Met

“…31 . To construct these scFvs, the heavy-chain (V H ) region was amplified using an upstream primer containing a SfiI site and a downstream primer encompassing the 3 0 end of the sequence.…”