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Development of HEK-293 Cell Lines Constitutively Expressing Flaviviral Antigens for Use in Diagnostics.
Powers, Jordan A; Skinner, Benjamin; Davis, Brent S; Biggerstaff, Brad J; Robb, Lucy; Gordon, Elizabeth; de Souza, William M; Fumagalli, Marcilio Jorge; Calvert, Amanda E; Chang, Gwong-Jen.
Afiliación
  • Powers JA; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Skinner B; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Davis BS; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Biggerstaff BJ; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Robb L; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Gordon E; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • de Souza WM; World Reference Center for Emerging Viruses and Arboviruses and Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA.
  • Fumagalli MJ; Centro de Pesquisa em Virologia, Faculdade de Medicina de Ribeirão Preto, Ribeirão Preto, Brazil.
  • Calvert AE; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
  • Chang GJ; U.S. Centers for Disease Control and Preventiongrid.416738.f, Fort Collins, Colorado, USA.
Microbiol Spectr ; 10(3): e0059222, 2022 06 29.
Article en En | MEDLINE | ID: mdl-35532242
Flaviviruses are important human pathogens worldwide. Diagnostic testing for these viruses is difficult because many of the pathogens require specialized biocontainment. To address this issue, we generated 39 virus-like particle (VLP)- and nonstructural protein 1 (NS1)-secreting stable cell lines in HEK-293 cells of 13 different flaviviruses, including dengue, yellow fever, Japanese encephalitis, West Nile, St. Louis encephalitis, Zika, Rocio, Ilheus, Usutu, and Powassan viruses. Antigen secretion was stable for at least 10 cell passages, as measured by enzyme-linked immunosorbent assays and immunofluorescence assays. Thirty-five cell lines (90%) had stable antigen expression over 10 passages, with three of these cell lines (7%) increasing in antigen expression and one cell line (3%) decreasing in antigen expression. Antigen secretion in the HEK-293 cell lines was higher than in previously developed COS-1 cell line counterparts. These antigens can replace current antigens derived from live or inactivated virus for safer use in diagnostic testing. IMPORTANCE Serological diagnostic testing for flaviviral infections is hindered by the need for specialized biocontainment for preparation of reagents and assay implementation. The use of previously developed COS-1 cell lines secreting noninfectious recombinant viral antigen is limited due to diminished antigen secretion over time. Here, we describe the generation of 39 flaviviral virus-like particle (VLP)- and nonstructural protein 1 (NS1)-secreting stable cell lines in HEK-293 cells representing 13 medically important flaviviruses. Antigen production was more stable and statistically higher in these newly developed cell lines than in their COS-1 cell line counterparts. The use of these cell lines for production of flaviviral antigens will expand serological diagnostic testing of flaviviruses worldwide.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Flavivirus / Flavivirus / Virus Zika / Infección por el Virus Zika Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Microbiol Spectr Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Flavivirus / Flavivirus / Virus Zika / Infección por el Virus Zika Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Microbiol Spectr Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos