Abstract
Objective: Evaluate the messenger RNAs (mRNA) PIWI proteins expression during infection with VDEN 4 in human hepatocyte cells. Materials and Methods: VDEN4 strain H778494 (JQ513335) was used, which was stored in Aedes albopictus cell culture (Clone C6 / 36) at the Arbovirology and Hemorrhagic Fevers section of the Evandro Chagas Institute. The techniques of cell cultures, stock (C6/36), inoculation, extraction, viral load quantification, RTqPCR and statistical analyzes were performed at the Viral Biogenesis Laboratory. Results: According to the results obtained, two cells (HepG2 and Huh7.5) demonstrated a higher level of viral replication at 72 hours post infection (hpi). The PIWI 2 target alter its mRNA expression during VDEN4 infection. The PIWI 4 target expression, was observed an altered expression in the infected cells. Thus, it was found that they are in different poles, while the viral load in the first three days showed high expression. The expression of mRNA was low in relation to the normal rate given by the uninfected cells. Conclusion: In our findings, it was observed that PIWI2 and 4 proteins have an inverse relationship to the viral infection process by VDEN4, when there is an increase in viral replication, these two proteins end up having a significant reduction in their expression. Probably, this reduction of expression is involved with the biogenesis process of apoptosis-regulating microRNAs.
Highlights
Viruses characterized by their association with arthropods, by their means of transmission, are described as arboviruses and stand out due their enzootic amplification and their impact at the epidemiological level, responsible for causing outbreaks in tropical centers, as if observed in Dengue, constituting a disease with a broad seasonal spectrum, associated with multifactorial dependence on determinants that influence its corresponding virus, such as temperature, humidity and climate (Amorim et al, 2020; Weaver, Reisen, 2010).The dengue virus (VDEN) is described as one of the most relevant arboviruses today, responsible for affecting human beings worldwide
It was observed that the PIWI targets 1 and 3 did not have significant changes between the cells infected by VDEN4 and the negative controls for both the HepG2 and Huh7.5 cell lines, the quantification of the PIWI targets 2 and 4 was possible to verify that they have differences in expression of their messenger RNAs (mRNA) in relation to uninfected control
The PIWI 2 mRNA target demonstrated that it occurs an expression changes during VDEN4 infection, being possible to relate a drop in its expression levels (Figure 2), and in the Huh 7.5 cell line a low expression of high significance was observed in to control between 24 to 72 hpi
Summary
Viruses characterized by their association with arthropods, by their means of transmission, are described as arboviruses and stand out due their enzootic amplification and their impact at the epidemiological level, responsible for causing outbreaks in tropical centers, as if observed in Dengue, constituting a disease with a broad seasonal spectrum, associated with multifactorial dependence on determinants that influence its corresponding virus, such as temperature, humidity and climate (Amorim et al, 2020; Weaver, Reisen, 2010). The dengue virus (VDEN) is described as one of the most relevant arboviruses today, responsible for affecting human beings worldwide. VDEN is divided into four serotypes VDEN1, VDEN2, VDEN3 and VDEN4, morphologically they are spherical viruses, measuring about 40 - 70 nm in diameter.
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