A long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice

Jennifer J. Brown, Bhupesh Parashar, Han Moshage, Kathryn E. Tanaka, Dean Engelhardt, Elazar Rabbani, Namita Roy-Chowdhury, Jayanta Roy-Chowdhury

Research output: Contribution to journalArticle

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Abstract

Development of new therapies for human hepatitis B virus infection (HBV) would be greatly facilitated by the availability of a suitable small-animal model for HBV virus production in vivo. To develop a murine model for HBV production, we established an immortalized, cloned liver cell line by transferring the Simian Virus 40 Large T-Antigen into primary human hepatocytes. These cells were stably transfected with a full-length HBV genome to generate a clone that expresses HBV genes and replicates HBV. The HBV-producing cells were transplanted into the livers of mice with combined immunodeficiency (Rag-2 deficient) by intrasplenic injection. Survival of the engrafted human hepatocytes was shown in several ways: fluorescent in situ hybridization (FISH) with a human-chromosome-specific DNA probe (human alpha satellite), dot-blot hybridization of the genomic DNA extracted from liver biopsy specimens with a human-specific Alu repetitive DNA probe, Blur-8, as well as with an HBV DNA probe, and secretion of human proteins into plasma. Histological examination of mouse liver up to 8 months following human cell transplant shows completely normal architecture. Determination of plasma HBV DNA levels indicated that engrafted cells secreted 3 x 107 to 3 x 108 virions per mL into the blood, and HBsAg was detected in plasma. This new murine model of HBV viremia should be useful for in vivo HBV studies.

Original languageEnglish (US)
Pages (from-to)173-181
Number of pages9
JournalHepatology
Volume31
Issue number1
StatePublished - 2000
Externally publishedYes

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Viremia
Virus Diseases
Hepatitis B
Hepatitis B virus
Hepatocytes
DNA Probes
Liver
Animal Hepatitis
Wiskott-Aldrich Syndrome
Nucleic Acid Hybridization
Simian virus 40
Viral Tumor Antigens
DNA
Human Chromosomes
Hepatitis B Surface Antigens
Fluorescence In Situ Hybridization
Virion
Blood Proteins
Animal Models
Clone Cells

ASJC Scopus subject areas

  • Hepatology

Cite this

A long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice. / Brown, Jennifer J.; Parashar, Bhupesh; Moshage, Han; Tanaka, Kathryn E.; Engelhardt, Dean; Rabbani, Elazar; Roy-Chowdhury, Namita; Roy-Chowdhury, Jayanta.

In: Hepatology, Vol. 31, No. 1, 2000, p. 173-181.

Research output: Contribution to journalArticle

Brown, JJ, Parashar, B, Moshage, H, Tanaka, KE, Engelhardt, D, Rabbani, E, Roy-Chowdhury, N & Roy-Chowdhury, J 2000, 'A long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice', Hepatology, vol. 31, no. 1, pp. 173-181.
Brown, Jennifer J. ; Parashar, Bhupesh ; Moshage, Han ; Tanaka, Kathryn E. ; Engelhardt, Dean ; Rabbani, Elazar ; Roy-Chowdhury, Namita ; Roy-Chowdhury, Jayanta. / A long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice. In: Hepatology. 2000 ; Vol. 31, No. 1. pp. 173-181.
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