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Using recombinant DNA technology for the development of live-attenuated dengue vaccines.

Institute of Biotechnology, National Tsing Hua University, Hsinchu 30013, Taiwan.
Enzyme and microbial technology 07/2012; 51(2):67-72. DOI: 10.1016/j.enzmictec.2012.05.005
Source: PubMed

ABSTRACT Dramatic increases in dengue (DEN) incidence and disease severity have been reported, in great part due to the geographic expansion of Aedes aegypti and Aedes albopictus mosquitoes. One result is the expanded co-circulation of all dengue 1-4 serotype viruses (DENV) in urban areas worldwide, especially in South and South-East Asia, and South America. DEN disease severity ranges from asymptomatic infections to febrile dengue fevers (DF) to life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). There is an urgent need for a safe and effective tetravalent DEN vaccine. Several live attenuated, tetravalent DEN vaccine candidates have been generated by recombinant DNA technology; these candidates are capable of providing immunity to all four DENV serotypes. In this paper we review (a) recombinant live-attenuated DEN vaccine candidates in terms of deletion, antigen chimerization, and the introduction of adaptive mutations; (b) strategies for improving tetravalent vaccine attenuation; and (c) live-attenuated DENV vaccine development.

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    ABSTRACT: Dengue virus belongs to Flavivirus and contains a positive-stranded RNA genome Binding of dengue virus to host cells was mediated through domain III of the viral envelope protein. Many therapeutic monoclonal antibodies (mAbs) against domain III have been generated and characterized because of its high antigenicity. We have previously established a novel PCR method named linear array epitope (LAE) technique for producing monoclone-like polyclonal antibodies. To prove this method could be utilized to produce antibody against epitopes with low antigenicity, a region of 10 amino acids (V365NIEAEPPFG374) from domain III of the envelope protein in dengue virus serotype 2 (DENV2) was selected to design the primers for LAE technique. A DNA fragment encoding 10 directed repeats of these 10 amino acids for producing the tandem repeated peptides was obtained and fused it with GST-containing vector. This fusion protein (GST-Den EIII10-His6) was purified from E. coli and used as antigen for immunizing rabbits to obtain polyclonal antibody. Furthermore, this EIII antibody could recognize envelope proteins either ectopically overexpressed or synthesized by DENV2 infection using immunoblot and immunofluorescence assays. Most importantly, this antibody was also capable of detecting DENV2 virions by ELISA assay and could block viral entry into BHK-21 cells as shown by immunofluorescence and qRT-PCR assays. Taken together, LAE technique could be applied for production of antibody against antigen with low antigenicity successfully and carries a high potential to produce antibodies with good quality for academic research, diagnosis and even therapeutic application in the future.
    Journal of General Virology 06/2014; · 3.53 Impact Factor

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