ABSTRACT
Arboviruses, including the Zika virus, have recently emerged as one of the most important threats to human health. The use of metagenomics-based approaches has already proven valuable to aid surveillance of arboviral infections, and the ability to reconstruct complete viral genomes from metatranscriptomics data is key to the development of new control strategies for these diseases. Herein, we used RNA-based metatranscriptomics associated with Ion Torrent deep sequencing to allow for the high-quality reconstitution of an outbreak-related Zika virus (ZIKV) genome (10,739 nt), with extended 5'-UTR and 3'-UTR regions, using a newly-implemented bioinformatics approach. Besides allowing for the assembly of one of the largest complete ZIKV genomes to date, our strategy also yielded high-quality complete genomes of two arthropod-infecting viruses co-infecting C6/36 cell lines, namely: Alphamesonivirus 1 strain Salvador (20,194 nt) and Aedes albopictus totivirus-like (4618 nt); the latter likely represents a new viral species. Altogether, our results demonstrate that our bioinformatics approach associated with Ion Torrent sequencing allows for the high-quality reconstruction of known and unknown viral genomes, overcoming the main limitation of RNA deep sequencing for virus identification.
Subject(s)
Arboviruses/genetics , Disease Outbreaks , Gene Expression Profiling/methods , Genome, Viral/genetics , High-Throughput Nucleotide Sequencing/methods , Nidovirales/genetics , Zika Virus Infection/virology , Zika Virus/genetics , Humans , Mosquito Vectors/virology , Phylogeny , Polymerase Chain Reaction , Zika Virus Infection/epidemiologyABSTRACT
The prevalence of allergic diseases in Brazil is one of the biggest in the world. Among these pathologies, we highlight asthma as one of the most importance. Asthma is characterized as a chronic inflammatory disease of airways, associated with hyperresponsiveness. Many environmental factors can trigger asthma symptoms, among them house dust mites can stimulate hypersensitivity type I reaction. The most common in house dust mite, in tropical countries, are Dermatophagoides pteronysinus and Blomia tropicalis. Several studies have shown that helminths, especially Schistosoma mansoni, lead to reduction of symptoms of atopy and allergic diseases. Therefore, the present study aims to evaluate the ability of recombinant S. mansoni proteins Sm200, and SmKI-1 to induce immunomodulation in vitro, using peripheral blood mononuclear cells (PBMCs) from atopic and non-atopic individuals, stimulated or not with B. tropicalis extract, and in vivo, in a murine model of allergy to the mite B. tropicalis. As results, we observed that the fragment called rSm200-3 and the protein rSmKI-1 stood out for their immunomodulatory potential, stimulating IL-10 production by human PBMCs in vitro. When these proteins were associated with B. tropicalis extract, it was observed the reduction of the production of the cytokine IL-5, with a statistically significant difference in non-atopic individual's cells. In vivo, both proteins presented similar results, with a reduction of IL-5 and IL-4 levels in lung homogenates and of serum IgE. SmKI-1 was also able to decrease the levels of EPO in lung homogenates and in BAL. These results showed that both proteins were able to downmodulate Th2 cells on human PBMCs, and in a murine model of allergy. However, SmKI-1 also reduced significantly the levels of EPO in BAL and lungs showing that this protein may be a good candidate to be used as a possible replacement or in conjunction with pharmacotherapy in individuals with unregulated immune response in asthma.
