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Surface Functionalization of Hepatitis E Virus Nanoparticles Using Chemical Conjugation Methods
Published on: May 11, 2018
[Construction and application of plasmid pVAX-HBVE harboring hepatitis B surface antibody targeted interferon and
Shasha Hou1, Yan Gao2, Yu Wang2
1College of Life Science, Qufu Normal University, Qufu 273165; Instiute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
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.
Conclusion:
The neo-type immune gene therapy plasmid was successfully constructed, which would provide an alternative for immune gene therapy of chronic hepatitis B.
