Antiviral activity and mechanism of action of arbidol against Hantaan virus infection

Purpose: To investigate the activity and mechanism of action of arbidol against Hantaan virus (HTNV) activity by modulating inflammation via TLR-4 pathway. Methods: HUVEC cells infected with HTNV 76-118 were treated with serially diluted arbidol solutions at -2h (2 h before viral infection, pre-treatment mode), 0 h (at the same time as viral infection, simultaneous treatment mode) or 2 h (2 h after viral infection, post-treatment mode). The transcript levels of TLR4 were detected by semi-quantitative reverse transcription-PCR (RT-PCR) at 6, 12, 18, and 24 h later. The levels of iNOS and TNF-α were examined using enzyme-linked immunosorbent assay (ELISA). Results: Pre-treatment with arbidol, rather than simultaneous treatment or post-treatment, effectively inhibited up-regulation of cellular TLR4 expression (up to 40 ± 6.1 % inhibition) and activity of supernatant iNOS induced by HTNV infection(up to 44.1 ± 9.4 % inhibition), as well as in a LPSstimulated inflammatory endothelial cell. Arbidol decreased the elevated TNF-α levels induced by LPS stimulation. Conclusion: These results are the first evidence that arbidol modulates viral PRRs signaling and its consequential inflammatory cytokine/chemokine response during hantavirus infection.

Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli

Background: Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Brazil, the Araraquara virus is one of the main causes of Hantavirus Cardio-Pulmonary Syndrome (HCPS). Methods: In this investigation, we report the expression of the N protein of the Araraquara Hantavirus in a Baculovirus Expression System, the use of this protein in IgM and IgG ELISA and comparison with the same antigen generated in E. coli. Results: The protein obtained, and purified in a nickel column, was effectively recognized by antibodies from confirmed HCPS patients. Comparison of the baculovirus generated antigen with the N protein produced in E. coli showed that both were equally effective in terms of sensitivity and specificity. Conclusions: Our results therefore indicate that either of these proteins can be used in serological tests in Brazil.

Anticorpos anti-hantavírus em escolares de Salvador, Bahia

As hantaviroses são doenças emergentes nas Américas e, em todo o mundo, os casos clínicos descritos foram infreqüentes entre crianças. O objetivo de investigar a freqüência de escolares portadores de anticorpos (IgG) anti-hantavírus justifica-se porque poucos estudos soroepidemiológicos pesquisaram a infecção por hantavírus no grupo pediátrico. Nos espécimes séricos de 379 escolares, de duas Escolas públicas da cidade do Salvador, Bahia, foram pesquisados os anticorpos anti-Hantaan (HTN) e anti-Sin Nombre (SN). A soropositividade de anticorpos anti-HTN foi de 13,2% (50/379) e todos os escolares foram soronegativos para o vírus SN. A soropositividade anti-HTN aumentou proporcionalmente com a idade. Não houve associação entre a soropositividade anti-HTN e as características relacionadas à exposição a roedores urbanos. A soroprevalência de anticorpos anti-HTN nos escolares estudados foi elevada e reforçou a hipótese de circulação de hantavírus em Salvador. A não observação de portadores de anticorpos anti-SN indica, provavelmente, a circulação de outros sorotipos nesta região, mais relacionados antigenicamente ao sorotipo Hantaan.