title: Evaluation of the cytopathicity (fusion/hemifusion) of patient-derived HIV-1 envelope glycoproteins comparing two effector cell lines.
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.
authors.affiliation: IrsiCaixa-HIVACAT, Institut de Recerca en Ciències de la Salut Germans Trias i Pujol (IGTP), Hospital Germans Trias, Universitat Autònoma de Barcelona, Barcelona, Catalonia, Spain.
authors.forename: Francesc
authors.initials: F
authors.lastname: Cunyat
authors.forename: Marta
authors.initials: M
authors.lastname: Curriu
authors.forename: Silvia
authors.initials: S
authors.lastname: Marfil
authors.forename: Elisabet
authors.initials: E
authors.lastname: García
authors.forename: Bonaventura
authors.initials: B
authors.lastname: Clotet
authors.forename: Julià
authors.initials: J
authors.lastname: Blanco
authors.forename: Cecilia
authors.initials: C
authors.lastname: Cabrera
mesh_terms: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002459:Cell Fusion; D003588:Cytopathogenic Effect, Viral; D005456:Fluorescent Dyes; D057809:HEK293 Cells; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015700:HIV Envelope Protein gp41 / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements; D008561:Membrane Fusion; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; C527276:CXCR4 protein, human; D005456:Fluorescent Dyes; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D019718:Receptors, CXCR4