Hepatitis B virus infection of tupaia hepatocytesin vitroandin vivo

Abstract

treatment of tree shrews in China 15,16 as well For the systematic analysis of various clinical and moas the easy adaptation of these animals to the laboratory lecular aspects of hepatitis B virus (HBV) infection, an environment, 17,18 we systematically analyzed the tree shrew experimental small animal system of HBV infection species tupaia belangeri for the study of HBV infection in would be a great advance. The susceptibility to HBV invitro and in vivo. fection, therefore, of hepatocytes from the tree shrew species tupaia belangeri was studied in vitro and in vivo. MATERIALS AND METHODS Primary hepatocytes isolated from livers of tupaias can be reproducibly infected with HBV. In vitro infectionTupaia Colony. For breeding purposes, animals (tupaia belangeri) were obtained from the German Primate Center, Göttingen, Gerresults in viral DNA and RNA synthesis in hepatocytes many. They were bred and maintained at the animal facilities of and secretion hepatitis B surface antigen (HBsAg) and the University Hospital Zü rich in accordance with institutionally hepatitis B e antigen (HBeAg) into culture medium. Tuapproved protocols. Under optimal conditions (room temperature, paias can also be infected with HBV in vivo, resulting 28ЊC; relative humidity, 70%; day-night light cycle, 12 and 12 hours), in viral DNA replication and gene expression in tupaia the mothers give birth to two to three animals every 8 to 10 weeks. livers. Similar to acute, self-limited hepatitis B in hu-At birth, the animals are about 6 to 7 cm long (without tail) and mans, HBsAg is rapidly cleared from serum, followed weigh 16 to 19 g. Within 10 to 12 weeks, they reach adulthood with by seroconversion to anti-HBe and anti-HBs. These data an average length of 12 to 18 cm (without tail) and 180 to 200 g body weight. All animals were negative for serological markers of HBV clearly show that HBV is infectious to tupaia hepato- problem worldwide and is associated with a wide spectrum of exclusion, was 90% to 95%. The cells were washed and seeded at a clinical presentations, ranging from the asymptomatic HBV density of 3 1 10 6 viable cells/10 mL medium on collagen-coated 100-carrier status to the development of cirrhosis and hepatocel-mm tissue culture plates. Plating efficiency was 85%. Cells were lular carcinoma. 1 HBV represents the prototypic member of maintained in Williams' E medium with Glutamax J, buffered with the hepadnaviruses 2,3 and many molecular and clinical as-15 mmol/L hydroxyethyl piperazine ethane sulfuric acid (Sigma), pH 7.4, and supplemented with 1.5% dimethyl sulfoxide (HybriMax, pects of HBV infection have been defined. 4,5 HBV naturally Sigma), 0.1 mmol/L insulin, 0.1 mmol/L dexamethasone, 100 mg/mL infects only humans and experimentally chimpanzees. While penicillin, and 100 mg/mL streptomycine.various aspects of the pathogenesis of HBV-related liver disTransient Transfection of Primary Tupaia Hepatocytes. Transfecease can be studied in transgenic animals 6-9 and animal modtion of primary tupaia hepatocytes with a replicatio...

Transmission of hepatitis C virus infection to tree shrews

“…assamensts (Wang et al, 1991). Tree shrews have shown susceptibility to infection by human rotavirus (Wan et al, 1982), herpes simplex virus type 1 and 2 (Darai et al, 1978), and human hepatitis viruses A, B, and D (Li et al, 1995;Walter et al, 1996;Yan et al, 1996a,b;Zan et al, 1981). Susceptibility of Tupaia to HCV infection has not been reported.…”

Adenoviral Delivery of Recombinant Hepatitis B Virus Expressing Foreign Antigenic Epitopes for Immunotherapy of Persistent Viral Infection

“…Huh-7 cells constitutively expressing the HBV core antigen were generated by lentiviral vector transduction. Primary tupaia hepatocytes (PTHs) were isolated by a twostep perfusion protocol (17,18). For viral infection, 1 ϫ 10 6 freshly isolated PTHs were seeded in collagen-coated 6-well plates (BD Biosciences, USA); 1 day after seeding, the PTHs were inoculated with wtHBV isolated from the infectious sera of HBV-infected patients or rHBV virions concentrated from culture supernatants by polyethylene glycol 8000.…”

Modeling hepatitis B virus infection, immunopathology and therapy in mice

“…In addition to higher primates, the tree shrew is the only other animal that can be experimentally infected with HBV (Walter et al, 1996; Yan et al, 1996). HBV infection in tree shrew results in viral replication in the liver, HBsAg secretion in the serum and subsequent appearance of anti-HBe and anti-HBs antibodies, recapitulating many aspects of self-limited acute hepatitis in humans (Walter et al, 1996).…”