25º Congresso Brasileiro de Microbiologia

25º Congresso Brasileiro de Microbiologia

ResumoID:754-1

Área: Virologia ( Divisão P )

CONSTRUCTION OF A RECOMBINANT MODIFIED VACCINIA ANKARA VIRUS EXPRESSING THE ENVELOPE PROTEIN OF DENGUE VIRUS SEROTYPE 2

Leandro Ciesielski Vida (UFMG); Tânia Mara Gomes Pinho (UFMG); Fabiana Magalhães Coelho (UFMG); Bárbara Resende Quinan (UFMG); Iara Apolinário Borges (UFMG); Danielle Daian (UFMG); Jaqueline Germano de Oliveira (CPqRR); Flávio Guimarães da Fonseca (UFMG)

Resumo

Introduction: Dengue virus (DENV) epidemics are a major public health problem in subtropical and tropical countries, such as Brazil. Despite decades of research, the development of a safe and effective vaccine against DENV is still on its experimental stages and consequently not available. The E protein, present in the envelope of the virus particle, is immunodominant and has the capacity to induce the production of neutralizing antibodies upon host infection. The protein also contains numerous T CD4+ and CD8+ lymphocytes recognition epitopes that present the potential to generate important cellular responses. The Modified Vaccinia virus Ankara (MVA) is a highly attenuated virus which is non-replicative in mammalian cells. An MVA vector, as opposed to inactive virus or recombinant protein vaccines, generates a significant cellular response, which may be a defining factor in the development of an effective vaccine against dengue infections. Objectives: to construct a recombinant MVA viral vector expressing the E protein of the DENV serotype 2 (DENV2). Methods: The recombinant viral vector was obtained by homologous recombination between a shuttle plasmid containing the cDNA encoding the E protein from DENV2 and the MVA genome. Cells previously infected with MVA were transfected with the shuttle plasmid (pLW44) containing the encoding gene for DENV2 E protein and recombinant viruses were picked on the basis of green fluorescence, conferred by a GFP gene also contained on the plasmid. Results and Discussion: The recombinant viruses have been generated and subjected to several successive selection rounds, followed by clone amplification and purification. The production of the recombinant DENV2E has been evaluated by methods including western blot and RT-PCR. The recombinant vector can now be tested as monovalent vaccine, so that, in the future, it may become a component in a tetravalent vaccine against DENV infection.਀䔀渀搀渀漀琀攀㨀吀栀椀猀瀀爀漀樀攀挀琀眀愀猀昀甀渀搀攀搀戀礀䘀䤀伀䌀刀唀娀Ⰰ 琀栀爀漀甀最栀琀栀攀倀䐀吀䤀匀瀀爀漀最爀愀洀Ⰰ 䌀䄀倀䔀匀Ⰰ 䌀一倀焀愀渀搀䘀䄀倀䔀䴀䤀䜀⸀

Palavras-chave: Dengue, MVA, Vaccine, E protein

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