title: [Establishment of an HBV chronic hepatitis B infection mouse model by vivo transduction of HBV cccDNA].
abstract: OBJECTIVE
  To generate a mouse model of chronic hepatitis B (CHB) infection by performing in vivo transduction of hepatitis B virus (HBV) covalently closed circular (ccc)DNA.
  
  
  METHODS
  Nude mice were injected with HBV cccDNA at doses of 1.5, 1.0 or 0.5 mug/ml. A control group was generated by giving equal injection volumes of physiological saline. The serum levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) on post-injection days 1 and 3, weeks 1-6, 8 and 10 were assayed by reflection immunoassay. At post-injection week 10, all animals were sacrificed and liver tissues were collected. Copies of HBV DNA in serum and liver tissue were detected by real-time PCR. HBV antigens in liver tissue were detected of by immunohistochemistry. Pathological analysis of liver tissue carried out with hematoxylin-eosin staining. Linear correlation of data was determined by statistical analysis.
  
  
  RESULTS
  HBsAg and HBeAg were detected in sera from all three groups of cccDNA-injected mice staring at post-injection day 1 and lasting through week 10. The levels of HBsAg over the 10-week period showed two patterns of increase-decrease;the lowest level was detected at week 4 and the highest level was detected at week 8. In contrast, the levels of HBeAg over the 10-week period showed three patterns of increase-decrease; the lower levels were detected at weeks 2 and 4 and the higher levels at weeks 3 and 6. HBV DNA copies in liver tissues showed a cccDNA dose-dependent descending trend over the 10-week study period (1.5 mug/ml:1.14E+07 ± 6.51E+06 copies/g, 1.0 mug/ml:9.81E+06 ± 9.32E+06 copies/g, and 0.5 mug/ml:3.72E+06 ± 2.35E+06 copies/g; Pearson's r =0.979). HBV DNA copies in sera showed the pattern of 1.0 mug/ml cccDNA more than 1.5 mug/ml cccDNA more than 0.5 mug/ml cccDNA, and in general were higher than those detected in the liver tissues. Liver tissues from all cccDNA-injected mice showed positive immunohistochemistry staining for both HBsAg and HBeAg. HE staining showed that the liver tissues of all cccDNA-injected mice had severe fatty and vacuolar degeneration and less obvious structure of liver lobules (compared to the liver tissues from control mice).
  
  
  CONCLUSIONS
  The CHB mouse model successfully established in this study by in vivo transduction of HBV cccDNA may represent a useful tool to study the pathogenic mechanisms and potential antiviral treatments of human CHB.
authors.affiliation: Laboratory Animal Center, Key Laboratory of Molecular Biology on Infection Diseases and Institute for Viral Hepatitis, Chongqing medical University, Chongqing 400016, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhao
authors.forename: Xiaosong
authors.initials: X
authors.lastname: Li
authors.forename: Wenwei
authors.initials: W
authors.lastname: Yin
authors.forename: Xuefei
authors.initials: X
authors.lastname: Cai
authors.forename: Wenlu
authors.initials: W
authors.lastname: Zhang
authors.forename: Feilan
authors.initials: F
authors.lastname: Chen
authors.forename: Guoqi
authors.initials: G
authors.lastname: Lai
authors.forename: Ailong
authors.initials: A
authors.lastname: Huang
mesh_terms: D000818:Animals; D004270:DNA, Circular / Q000008:administration & dosage*; D004279:DNA, Viral / Q000008:administration & dosage*; D004195:Disease Models, Animal; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006513:Hepatitis B e Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology; D019694:Hepatitis B, Chronic / Q000821:virology*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D014161:Transduction, Genetic; D014779:Virus Replication
chemical_list: D004270:DNA, Circular; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens