New insights into HCV replication in original cells from Aedes mosquitoes

Abstract

BackgroundThe existing literature about HCV association with, and replication in mosquitoes is extremely poor. To fill this gap, we performed cellular investigations aimed at exploring (i) the capacity of HCV E1E2 glycoproteins to bind on Aedes mosquito cells and (ii) the ability of HCV serum particles (HCVsp) to replicate in these cell lines.MethodsFirst, we used purified E1E2 expressing baculovirus-derived HCV pseudo particles (bacHCVpp) so we could investigate their association with mosquito cell lines from Aedes aegypti (Aag-2) and Aedes albopictus (C6/36). We initiated a series of infections of both mosquito cells (Ae aegypti and Ae albopictus) with the HCVsp (Lat strain - genotype 3) and we observed the evolution dynamics of viral populations within cells over the course of infection via next-generation sequencing (NGS) experiments.ResultsOur binding assays revealed bacHCVpp an association with the mosquito cells, at comparable levels obtained with human hepatocytes (HepaRG cells) used as a control. In our infection experiments, the HCV RNA (+) were detectable by RT-PCR in the cells between 21 and 28 days post-infection (p.i.). In human hepatocytes HepaRG and Ae aegypti insect cells, NGS experiments revealed an increase of global viral diversity with a selection for a quasi-species, suggesting a structuration of the population with elimination of deleterious mutations. The evolutionary pattern in Ae albopictus insect cells is different (stability of viral diversity and polymorphism).ConclusionsThese results demonstrate for the first time that natural HCV could really replicate within Aedes mosquitoes, a discovery which may have major consequences for public health as well as in vaccine development.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-017-0828-z) contains supplementary material, which is available to authorized users.

Genome Wide Mapping of Peptidases in Rhodnius prolixus: Identification of Protease Gene Duplications, Horizontally Transferred Proteases and Analysis of Peptidase A1 Structures, with Considerations on Their Role in the Evolution of Hematophagy in Triatominae

“…Hepatitis virus has been found to be stable and viable for at least 1 month in the gut of bed bugs ( Cymex lectularius and C. hemipterus ) and R. prolixus , with the virus being detected in feces and transmitted trans stadially (Blow et al, 2001 ; Silverman et al, 2001 ). Besides that, Hepatitis virus has been detected in mosquitoes in endemic areas, and it has been found that Hepatitis virus efficiently replicates in cultures of mosquito cells (Fallecker et al, 2017 ). The possibility of Hepatitis transmission by arthopods has been hindered by the ignorance of natural reservoirs of the virus or documentation of prevalence of the virus in putative vectors (Houldsworth, 2017 ).…”

Examining the Role of Niemann–Pick C1 Protein in the Permissiveness of Aedes Mosquitoes to Filoviruses

“…The mosquito is the most profound vector of disease in the world . While several flaviviruses such as dengue and Zika are transmitted to humans by the Aedes mosquito, hepatitis C virus is a human flavivirus with no arthropod vector. , The factors that distinguish viruses capable of being vectored by insects as well as the capacity for viruses to gain this ability have not been fully elucidated and remain unclear. In summary, we have shown that the expression of human NPC1 protein in Aedes cells alone is not sufficient to permit filovirus infection, indicating at the presence of multiple blocks to infection.…”

“…Many significant arboviruses, including Yellow Fever virus, West Nile virus, dengue virus, and Zika virus, belong to the family Flaviviridae and are vectored by mosquitoes belonging to the genus Aedes . However, certain other members of the Flaviviridae family, such as hepatitis C virus, with similar genomes and structures to these arboviruses, are not vectored by the mosquito. , The molecular mechanisms responsible for this distinction within the Flaviviridae family are unknown but bring up the questions of what factors may be necessary for a virus to be vectored by an arthropod and which viruses could potentially gain this ability, especially among closely related viruses.…”

The MultiBac system: a perspective

“…MultiBac is successfully used for a range of applications beyond producing protein complexes for structural and mechanistic studies. For example, customized MultiBac baculoviral genomes were prepared to express virus-like 15 particles (VLPs), which are promising vaccine candidates as they resemble live viruses but do not contain genetic material and are thus safe [22,39]. Here, an influenza VLP is shown in a schematic drawing and in an electron micrograph [22] (centre, top).…”