Análise da coinfecção de Dengue virus e Zika virus e do efeito dos inibidores farmacológicos de MEK/ERK durante a coinfecção

Abstract

Dengue is the most important arthropod-borne viral infection, and it is estimated that that 100 million are infected every year, with mortality rate close to 5% in the absence of treatment, and 26% to severe dengue. Dengue can be caused by any of the four distinct serotypes (DENV1-4). Zika virus caused, between 2015 and 2016, epidemics in America, Africa, Pacific and southeast of Asia, and has been declared a world public health concern by WHO due the disease association with neurological problems and severe new born sequelae as microcephaly, reported in Brazil. Cocirculation of Dengue and other arboviruses such as Zika virus, yellow fever virus and Chikungunya virus have been frequently occurring on many endemic regions in Brazil. In this context, antivirals prospection must be evaluated on models that allow us to understand how the antiviral activity works during simultaneous infections. Initial coinfection analysis of DENV in VERO cell line showed that only DENV-1 was affected when coinfected with DENV-3 and DENV-4, with its genomic copy numbers reduced around 2 log10. No viral interference was observed between DENV-3 and DENV-4, and they were chosen to proceed with the coinfection experiments. Different MOIs did not change the results, as the superinfection with time lapse of 6 hours, suggesting that the virus is capable of infect a previous infected cell by another different virus. We compared the number of genomic copies with the number of viable particles (PFU/mL) and found a rate of 1:10000. We also did the multiplication curve of DENV-3 and DENV-4 at MOI 1, and no significative difference was observed between the samples. Coinfection between ZIKV and DENV-4 showed that DENV-4 had its genomic copies reduced around 1 log10 in 24, 48 and 72 h.p.i. The use of the inhibitors MEKi-A and MEKi-B, had no effect in replication of DENV-3 and DENV-4, regardless of coinfection.