Experimental Study On Immunotherapy Of Chronic Hepatitis B Virus Gene

Antiviral therapy of hepatitis B treatment is the most effective means, nucleoside analoguesand interferon for replication and can inhibit the synthesis of hepatitis B virus and is often usedin clinical antiviral therapy, interferon also has a role in immune regulation. Although antiviraldrugs commonly used can effectively reduce the amount of virus in patients with the body, andsome even can be reduced to undetectable.But soon after the withdrawal will be repeated illness,long-term medication nucleoside drugs prone to drug resistance, interferon treatment regimenlong, expensive and so on. Exploring new effective treatment has become a pressing matter ofthe moment for the treatment of chronic hepatitis B.Gene therapy (gene therapy) refers to the antiviral factor into target cells by exogenous,cytokine gene and expression of costimulatory molecule gene suppression of viral replication indifferent level, effectively induce immune responses, but also avoid the resistance caused bylong-term administration and side effects. As a new treatment method can effectively avoid thecause drug resistance and side effects, application prospect of gene therapy is very broad.Choose the appropriate carrier, the objective gene into target cells, the target cells in genetherapy is decided the success or failure of important one annulus. Current the most widely usedclinical gene therapy agent is based on viral vector,which has the advantages of virus infectedcells ability, immune gene into the target cell, gene therapy effect is obvious; but its drawback isthat its immunogenicity of viral vector is very strong and can not be repeatedly used, otherwisethe body will generate immune response against the carrier own, it will offset the effect of genetherapy. Plasmid vectors for gene therapy agent has the advantages of high safety, lowproduction cost and good stability and so on. At the same time, plasmid vector has large capacity,it can easily realize the objective gene combination. Compared to viral vectors, plasmid vectorhas its low immunogenicity, and can be repeatedly used, without triggering the body’s immuneresponse against the carrier. Although delivery efficiency of the plasmid into cell is lower thanviral vectors, but with the development of scientific research, plasmid DNA delivery technologyhas achieved great development, especially the application of electroporation technology, makesthe plasmid delivered into cells and increase the efficiency of a hundred times, efficiency ofinfection are almost close to those viral vectors.This study constructed fusion expression of human interferon alpha and interleukin-12immune gene therapy plasmid pVAX-HBVE and replicating plasmid pSVK-HBVE, plasmid issuccessfully constructed the enzyme digestion and sequencing showed that, to obtain secreted expression in the corresponding cell lines, through the ELISA to detect the expression efficiencyof plasmid, and the use of a transgenic mouse model of copy the immune gene therapy plasmidpVAX-HBVE and pSVK-HBVE was primarily immune function verification, and the immunemechanism of in-depth study, laid the foundation for the treatment of immune function ofcomprehensive validation of plasmid...