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Evaluation of interaction between neutrophils extracellular traps (NETs) and Human respiratory syncicial Virus (hRSV)


Respiratory Syncytial Virus (hRSV) is the major etiologic agent of acute lower respiratory tract infections, resulting in more than 3 million very severe cases of bronchiolitis throughout the year. During infection, leukocytes are massively recruited to the infectious site where they secrete cytokines and other inflammatory mediators. About 70% of these cells are neutrophils that, in addition to secreting the contents of their granules, can secrete extracellular structures that form a kind of microbicidal network, the NETs. These structures are mainly composed of DNA and granular / nuclear / cytoplasmic proteins and have the function of capturing and inactivating different types of microorganisms. Recent studies demonstrate that hRSV is able to induce the formation of NETs through its F protein, confirming clinical findings in the lungs of patients infected with hRSV. The excess of NETs, cell debris and mucus resulting from hRSV infection form a consistent mucus of difficult expectoration that accumulates in the lumen of the bronchioles, drastically restricting the flow of air, worsening the clinical picture of the patient. NETs have been shown to interact with the viral particles of hRSV, and recent data from our group indicate that this contact results in virucidal action of these structures.This study did not confirm the identity of which molecule (s) would be involved directly in NET virucidal effect. Thus, the objective of this work is to identify which NET molecule (s) would be responsible for that virucidal effect seen in the previous study. Immunobiological tests, both in silico and in vitro, will be carried out. To investigate the interaction between some macromolecules of the NETs and the viral protein F will be carried out analyzes, electrophoresis, Western Blot, docking / molecular dynamics and proteomics. The biological consequences of this interaction will be assessed by in vitro (virucidal) assays in HEp-2 cells using cultured lines and clinical samples of RSV. The data generated in this study could guide new strategies in the treatment of patients with hRSV, since the improvement in respiratory quality and in the blockade of viral dissemination. (AU)

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