title: [Factors influencing long-term hepatitis B virus infection of the tree shrew (Tupaia belangeri chinensis) as an in vivo model of chronic hepatitis B].
abstract: OBJECTIVE
  To determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).
  
  
  METHODS
  Seventy-seven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum, single or pooled; HBV-infected tree shrew serum, single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen). Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy.
  
  
  RESULTS
  Among the neonatal group of tree shrews, six (7.8%) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1%) were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.
  
  
  CONCLUSIONS
  In order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.
authors.affiliation: Department of Experimental Pathology, Guangxi Cancer Institute, Nanning 530021, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.forename: Chun
authors.initials: C
authors.lastname: Yang
authors.forename: Jian-jia
authors.initials: JJ
authors.lastname: Su
authors.forename: Ji
authors.initials: J
authors.lastname: Cao
authors.forename: Chao
authors.initials: C
authors.lastname: Ou
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: Jing-jing
authors.initials: JJ
authors.lastname: Zhang
authors.forename: Jun-lin
authors.initials: JL
authors.lastname: Shi
authors.forename: Dou-ping
authors.initials: DP
authors.lastname: Wang
authors.forename: Xiao-juan
authors.initials: XJ
authors.lastname: Wang
authors.forename: Jia
authors.initials: J
authors.lastname: Wan
authors.forename: Ping
authors.initials: P
authors.lastname: Ruan
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D056945:Hep G2 Cells; D006515:Hepatitis B virus; D019694:Hepatitis B, Chronic / Q000821:virology*; D006801:Humans; D008297:Male; D014417:Tupaia