title: Mutations in both env and gag genes are required for HIV-1 resistance to the polysulfonic dendrimer SPL2923, as corroborated by chimeric virus technology.
abstract: A drug-resistant NL4.3/SPL2923 strain has previously been generated by in vitro selection of HIV-1(NL4.3) in the presence of the polysulfonic dendrimer SPL2923 and mutations were reported in its gp120 gene (Witvrouw et al., 2000). Here, we further analysed the (cross) resistance profile of NL4.3/SPL2923. NL4.3/SPL2923 was found to contain additional mutations in gp41 and showed reduced susceptibility to SPL2923, dextran sulfate (DS) and enfuvirtide. To delineate to what extent the mutations in each env gene were accountable for the phenotypic (cross) resistance of NL4.3/SPL2923, the gp120-, gp41- and gp160-sequences derived from this strain were placed into a wild-type background using env chimeric virus technology (CVT). The cross resistance of NL4.3/SPL2923 towards DS was fully reproduced following gp160-recombination, while it was only partially reproduced following gp120- or gp41-recombination. The mutations in gp41 of NL4.3/SPL2923 were sufficient to reproduce the cross resistance to enfuvirtide. Unexpectedly, the reduced sensitivity towards SPL2923 was not fully reproduced after gp160-recombination. The search for mutations in NL4.3/SPL2923 in viral genes other than env revealed several mutations in the gene encoding the HIV p17 matrix protein (MA) and one mutation in the gene encoding the p24 capsid protein (CA). In order to analyse the impact of the gag mutations alone and in combination with the mutations in env on the phenotypic resistance towards SPL2923, we developed a novel p17- and p17/gp160-CVT. Phenotypic analysis of the NL4.3/SPL2923 p17- and p17/gp160-recombined strains indicated that the mutations in both env and gag have to be present to fully reproduce the resistance of NL4.3/SPL2923 towards SPL2923.
authors.affiliation: Laboratory for Molecular Virology, Molecular Medicine, Katholieke Universiteit Leuven, Flanders, Belgium.
authors.forename: Anke
authors.initials: A
authors.lastname: Hantson
authors.forename: Valery
authors.initials: V
authors.lastname: Fikkert
authors.forename: Barbara
authors.initials: B
authors.lastname: Van Remoortel
authors.forename: Chistophe
authors.initials: C
authors.lastname: Pannecouque
authors.forename: Peter
authors.initials: P
authors.lastname: Cherepanov
authors.forename: Barry
authors.initials: B
authors.lastname: Matthews
authors.forename: George
authors.initials: G
authors.lastname: Holan
authors.forename: Erik
authors.initials: E
authors.lastname: De Clercq
authors.forename: Anne-Mieke
authors.initials: AM
authors.lastname: Vandamme
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
authors.forename: Myriam
authors.initials: M
authors.lastname: Witvrouw
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D002460:Cell Line; D050091:Dendrimers / Q000494:pharmacology*; D024882:Drug Resistance, Viral / Q000235:genetics*; D015752:Genes, env / Q000235:genetics*; D015750:Genes, gag / Q000235:genetics*; D005818:Genetic Engineering; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D009154:Mutation; D010641:Phenotype; D011995:Recombination, Genetic; D013451:Sulfonic Acids / Q000494:pharmacology*
chemical_list: D019380:Anti-HIV Agents; C415374:BRI 2923; D050091:Dendrimers; D013451:Sulfonic Acids