Evaluation of the cytopathicity (fusion/hemifusion) of patient-derived HIV-1 envelope glycoproteins comparing two effector cell lines

Francesc Cunyat, Marta Curriu, Silvia Marfil, Elisabet García, Bonaventura Clotet, Julià Blanco, Cecilia Cabrera

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6 Citations (Scopus)

Abstract

HIV-1 envelope glycoprotein (Env) is a major determinant of viral pathogenicity. The evaluation of the biological properties of patient-derived envelopes by comparing two effector cell lines (293T and HeLa) is reported. A standard cell-to-cell fusion assay was used to evaluate fusogenicity, whereas a coculture with CD4+ cells was used to evaluate absolute cell loss, single cell death, and hemifusion events. Fusion and absolute cell loss assays showed that Env-expressing 293T and HeLa cells had different fusion efficiencies; fusion was magnified in 293T cells despite a significantly lower cell-surface Env expression. Conversely, gp41-mediated single cell death and hemifusion induced in CD4+ cells by 293T-Env-positive cells were significantly lower than that induced by HeLa-Env-positive cells. These data showed that the effector cell line used in the in vitro assays is crucial, and a combination of assays is recommended to evaluate the biological properties of patient-derived envelope glycoproteins: preferentially, 293T-Env-positive cells for the evaluation of fusogenicity and HeLa-Env-positive cells for the evaluation of cell death parameters. The combination of assays described in our work could be a valuable tool for dual screenings of large collections of primary Envs or Env mutants and drugs acting on these Envs. © 2012 Society for Laboratory Automation and Screening.
Original languageEnglish
Pages (from-to)727-737
JournalJournal of Biomolecular Screening
Volume17
Issue number6
DOIs
Publication statusPublished - 1 Jul 2012

Keywords

  • apoptosis
  • cytopathicity
  • envelope
  • fusion
  • hemifusion
  • HIV

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