title: [Establishing a L02 cell model with whole HBV gene transfection and analyzing its expressions of HLA-A, B, C and MICA/B].
abstract: OBJECTIVE
  To establish a cell model with HBV secretion by plasmid transfection with whole HBV genes into hepatic L02 cells, and to analyze the effect of HBV on the expression of HLA-A, B, C and MICA/B in L02 cells.
  
  
  METHODS
  The mock control plasmid was built by digesting pEcob6 plasmid with EcoR I at the HBV DNA site and ligating the fragment without HBV DNA. The L02 cells were transfected with pEcob6 or mock plasmid and pcDNA3 1-neo by liposome. The expressions of HBsAg, HBcAg/HBeAg and HBV DNA were detected by immunofluorescence assay, Abbott enzymoimmunoassay, or FQ-PCR. The expressions of HLA-A, B, C and MICA/B were determined by FACS and the differences in the two molecules were analyzed.
  
  
  RESULTS
  After the transfected cells were selected by G418, the HBsAg and HBeAg in the supernatant of L02-HBV cells were 24.78(S/N) and 4.117(S/N). The quantity of HBV DNA was 9.67 x 10(4) copies/ml, but in L02-mock and in L02 cells they were all negative. Under a confocal microscope HBsAg was brightly shown in the cytoplasm while HBcAg showed dimly in the cytoplasm or in the nuclei. By using FACS, the L02 and L02-mock cells showed some low expressions of HLA-A, B, C and a little expression of MICA/B, while the expression of the two molecules was higher in L02-HBV and the differences were significant (P < 0.05).
  
  
  CONCLUSIONS
  A cell model with the expression of HBV antigens and the secretion of HBV DNA was established by pEcob6 transfection into L02 cells, the transcription or replication of HBV gene might induce stronger expressions of HLA-A, B, C and MICA/B on hepatic cells. They might be related to the immune injuries of hepatic cells.
authors.affiliation: Key Laboratory of Molecular Biology for Infectious Diseases, PRC Ministry of Education, Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.forename: Ming-li
authors.initials: ML
authors.lastname: Peng
authors.forename: Qing-feng
authors.initials: QF
authors.lastname: Shi
authors.forename: Hong
authors.initials: H
authors.lastname: Ren
mesh_terms: D002460:Cell Line; D004279:DNA, Viral / Q000378:metabolism; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006680:HLA Antigens / Q000235:genetics*; D006511:Hepatitis B Antigens / Q000235:genetics / Q000378:metabolism*; D006515:Hepatitis B virus / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D014162:Transfection
chemical_list: D004279:DNA, Viral; D006680:HLA Antigens; D006511:Hepatitis B Antigens; D015395:Histocompatibility Antigens Class I; C099187:MHC class I-related chain A; C098080:MICB antigen