Human Brain Microvascular Endothelial Cells Exposure to SARS-CoV-2 Leads to Inflammatory Activation through NF-κB Non-Canonical Pathway and Mitochondrial Remodeling

Neurological effects of COVID-19 and long-COVID-19, as well as neuroinvasion by SARS-CoV-2, still pose several questions and are of both clinical and scientific relevance. We described the cellular and molecular effects of the human brain microvascular endothelial cells (HBMECs) in vitro exposure by SARS-CoV-2 to understand the underlying mechanisms of viral transmigration through the blood-brain barrier. Despite the low to non-productive viral replication, SARS-CoV-2-exposed cultures displayed increased immunoreactivity for cleaved caspase-3, an indicator of apoptotic cell death, tight junction protein expression, and immunolocalization. Transcriptomic profiling of SARS-CoV-2-challenged cultures revealed endothelial activation via NF-κB non-canonical pathway, including RELB overexpression and mitochondrial dysfunction. Additionally, SARS-CoV-2 led to altered secretion of key angiogenic factors and to significant changes in mitochondrial dynamics, with increased mitofusin-2 expression and increased mitochondrial networks. Endothelial activation and remodeling can further contribute to neuroinflammatory processes and lead to further BBB permeability in COVID-19

Low prevalence of influenza A strains with resistance markers in Brazil during 2017–2019 seasons

This project was supported by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Programa Estratégico de Apoio à Pesquisa em Saúde (PAPES), Fundação Oswaldo Cruz, CNPq, and Coordenação Geral de Laboratórios de Saúde Pública (CGLAB) from the Brazilian Ministry of Health.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Laboratório Central de Saúde Pública de Sergipe. Aracaju, SE, Brazil.Laboratório Central de Saúde Pública de Sergipe. Aracaju, SE, Brazil.Laboratório Central do Estado do Paraná. Curitiba, PR, Brazil.Laboratório Central do Estado do Paraná. Curitiba, PR, Brazil.Secretaria de Saúde do Estado do Espírito Santo. Laboratório de Saúde Pública do Estado do Espírito Santo. Vitória, ES, Brazil / Universidade Federal do Espírito Santo. Núcleo de Doenças Infecciosas. Vitória, ES, Brazil.Secretaria de Saúde do Estado do Espírito Santo. Laboratório de Saúde Pública do Estado do Espírito Santo. Vitória, ES, Brazil / Universidade Federal do Espírito Santo. Núcleo de Doenças Infecciosas. Vitória, ES, Brazil.Laboratório Central de Saúde Pública do Rio de Janeiro. Rio de Janeiro, RJ, Brazil.Laboratório Central de Saúde Pública do Rio de Janeiro. Rio de Janeiro, RJ, Brazil.Secretaria de Saúde do estado do Rio Grande do Sul. Laboratório Central de Saúde Pública. Porto Alegre, RS, Brazil.Secretaria de Saúde do estado do Rio Grande do Sul. Laboratório Central de Saúde Pública. Porto Alegre, RS, Brazil.Fundação Ezequiel Dias. Laboratório Central de Saúde Pública de Minas Gerais. Belo Horizonte, MG, Brazil.Fundação Ezequiel Dias. Laboratório Central de Saúde Pública de Minas Gerais. Belo Horizonte, MG, Brazil.Laboratório Central da Saúde Pública do estado da Bahia. Salvador, BA, Brazil.Laboratório Central da Saúde Pública do estado da Bahia. Salvador, BA, Brazil.Laboratório Central de Santa Catarina. Florianópolis, SC, Brazil.Laboratório Central de Santa Catarina. Florianópolis, SC, Brazil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Ananindeua, PA. Brasil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Ananindeua, PA. Brasil.Instituto Adolfo Lutz. Laboratório Central de Saúde Pública do Estado de São Paulo. São Paulo, SP, Brazil.Instituto Adolfo Lutz. Laboratório Central de Saúde Pública do Estado de São Paulo. São Paulo, SP, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de Imunização e Doenças Transmissíveis. Brasília, DF, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de Imunização e Doenças Transmissíveis. Brasília, DF, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.Fiocruz Fundation. Oswaldo Cruz Institute. Laboratory of Respiratory Viruses and Measles. Rio de Janeiro, RJ, Brazil.The influenza A virus (IAV) is of a major public health concern as it causes annual epidemics and has the potential to cause pandemics. At present, the neuraminidase inhibitors (NAIs) are the most widely used anti-influenza drugs, but, more recently, the drug baloxavir marboxil (BXM), a polymerase inhibitor, has also been licensed in some countries. Mutations in the viral genes that encode the antiviral targets can lead to treatment resistance. Worldwide, a low prevalence of antiviral resistant strains has been reported. Despite that, this situation can change rapidly, and resistant strain surveillance is a priority. Thus, the aim of this was to evaluate Brazilian IAVs antiviral resistance from 2017 to 2019 through the identification of viral mutations associated with reduced inhibition of the drugs and by testing the susceptibility of IAV isolates to oseltamivir (OST), the most widely used NAI drug in the country. Initially, we analyzed 282 influenza A(H1N1)pdm09 and 455 A(H3N2) genetic sequences available on GISAID. The amino acid substitution (AAS) NA:S247N was detected in one A(H1N1)pdm09 strain. We also identified NA:I222V (n = 6) and NA:N329K (n = 1) in A(H3N2) strains. In addition, we performed a molecular screening for NA:H275Y in 437 A(H1N1)pdm09 samples, by pyrosequencing, which revealed a single virus harboring this mutation. Furthermore, the determination of OST IC50 values for 222 A(H1N1)pdm09 and 83 A(H3N2) isolates revealed that all isolates presented a normal susceptibility profile to the drug. Interestingly, we detected one A(H3N2) virus presenting with PA:E119D AAS. Moreover, the majority of the IAV sequences had the M2:S31N adamantanes resistant marker. In conclusion, we show a low prevalence of Brazilian IAV strains with NAI resistance markers, in accordance with what is reported worldwide, indicating that NAIs still remain an option for the treatment of influenza infections in Brazil. However, surveillance of influenza resistance should be strengthened in the country for improving the representativeness of investigated viruses and the robustness of the analysis