title: [Construction and application of plasmid pVAX-HBVE harboring hepatitis B surface antibody targeted interferon and interleukin 12 genes].
abstract: OBJECTIVE
  To construct a new gene therapy plasmid that can express both hepatitis B surface antibody(HBsAb) targeted interferon (dsFvα) and human interleukin 12 (hIL-12) genes for the immunotherapy of chronic hepatitis B.
  
  
  METHODS
  The pEE14.1-dsFvα plasmid was digested to obtain dsFvα fragment, and then the fragment was cloned into the upstream of IRES sequence in vector pVAX-IRES-hIL-12 digested by the same enzyme to construct the recombinant expression plasmid pVAX-HBVE. The recombinant plasmid was transiently transfected into the HEK293T cells and the expression of target gene was detected by ELISA. The recombinant plasmid pVAX-HBVE was extracted and injected into the leg muscles of HBV transgenic mice with electroporation delivery. The HBV gene copies were detected by quantitative PCR before and after injection.
  
  
  RESULTS
  Enzyme digestion and sequencing analysis showed that the recombinant plasmid pVAX-HBVE was constructed as expected before. ELISA showed that dsFvα gene and IL-12 gene were successfully expressed in the supernatant of transfected cells. The recombinant plasmid pVAX-HBVE (30 μg, once) reduced the HBV gene copy by two orders of magnitude after being injected into transgenic mice.
  
  
  CONCLUSIONS
  The neo-type immune gene therapy plasmid was successfully constructed, which would provide an alternative for immune gene therapy of chronic hepatitis B.
authors.affiliation: College of Life Science, Qufu Normal University, Qufu 273165; Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Shasha
authors.initials: S
authors.lastname: Hou
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wu
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Zhu
authors.affiliation: College of Life Science, Qufu Normal University, Qufu 273165, China.
authors.forename: Bingran
authors.initials: B
authors.lastname: Guo
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yuanji
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Jinqi
authors.initials: J
authors.lastname: Yan
authors.affiliation: Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Jiyun
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D017077:Culture Media, Conditioned / Q000378:metabolism; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D057809:HEK293 Cells; D006509:Hepatitis B / Q000097:blood / Q000276:immunology* / Q000628:therapy; D006510:Hepatitis B Antibodies / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007372:Interferons / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018664:Interleukin-12 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008822:Mice, Transgenic; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics*; D014162:Transfection
chemical_list: D017077:Culture Media, Conditioned; D006510:Hepatitis B Antibodies; D018664:Interleukin-12; D007372:Interferons