VSV-G pseudotyping rescues HIV-1 CA mutations that impair core assembly or stability

Sonia Brun¹^,2^,3 , Maxime Solignat¹^,2^,3 , Bernard Gay¹^,2^,3 , Eric Bernard¹^,2^,3 , Laurent Chaloin¹^,2^,3 , David Fenard¹^,2^,3^,4 , Christian Devaux¹^,2^,3 , Nathalie Chazal¹^,2^,3 and Laurence Briant¹^,2^,3

¹Université Montpellier 1, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), France

²CNRS, UMR 5236, CPBS, F-34965, Montpellier, France

³Université Montpellier 2, CPBS, F-34095, Montpellier, France

⁴GENETHON, 1bis rue de l'Internationale – BP60, 91002 EVRY cedex, France

author email corresponding author email

Retrovirology 2008, 5:57doi:10.1186/1742-4690-5-57


Published:	7 July 2008

Additional files

Additional file 1:

Infectivity of increasing amounts of WT viruses and CA mutants expressing HIV Env or VSV-G. Viral inputs were normalized according to RTase activity (ranging from 100 to 10,000 cpm) and used to infect MAGIC-5B cells. 48 hours post-infection, relative infectivity was measured by quantification of o-nitrophenyl β-D-galactopyranoside hydrolysis. Time of incubation used for revelation of β-galactosidase assay was adapted to produce non saturating OD at 405 nm (1h45 in panel A and 25 min in panel B). For each dose tested, infectivity of S₁₄₉A and S₁₇₈A mutants is indicated as a percentage of β-galactosidase activity generated by an identical amount of WT viruses with the corresponding envelope.

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Additional file 2:

Quantification of β-galactosidase activities generated in MAGIC-5B cells infected with amounts of CA mutants expressing HIV Env or VSV-G that generated comparable strong-stop DNA copy numbers. MAGIC-5B cells were infected with S₁₄₉A or S₁₇₈A mutants expressing HIV Env or VSV-G, using infectious doses that generated similar strong-stop DNA copy numbers as measured by qPCR 24 h post-infection. Relative infectivity is expressed as o-nitrophenyl β-D-galactopyranoside hydrolysis measured from total cell extracts by absorbance at 405 nm. Each value represents an average of two experiments performed in duplicate ± standard deviation.

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