Abstract
Several small molecule drugs that bind to the host CCR5 co-receptor and prevent viral entry have been developed for the treatment of HIV-1 infection. The innate variability found in HIV-1 envelope and the complex viral/cellular interactions during entry makes defining resistance to these inhibitors challenging. Here we found that mapping determinants in the gp160 gene from a primary isolate RU570-VCV(res), selected in culture for resistance to the CCR5 entry inhibitor vicriviroc, was complicated by inactivity of the cloned envelope gene in pseudovirus assays. We therefore recombined the envelope from RU570-VCV(res) into a highly active and susceptible ADA gp160 backbone. The chimeric envelopes generated robust signals in the pseudovirus assay and a 200 amino acid fragment, encompassing a C2-V5 region of the RU570-VCV(res) envelope, was required to confer resistance in both the single-cycle assay and in replicating virus. In contrast, a chimeric envelope that contained only the V3-loop region from this resistant virus was completely susceptible suggesting that the V3-loop changes acquired are context dependent.
MeSH terms
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Anti-HIV Agents / pharmacology
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Base Sequence
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CCR5 Receptor Antagonists*
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Cell Line
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Chimera
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Chromosome Mapping
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DNA Primers / genetics
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DNA, Viral / genetics
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Drug Resistance, Viral / genetics
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Genes, env*
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HIV Envelope Protein gp120 / chemistry
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HIV Envelope Protein gp120 / genetics*
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HIV Envelope Protein gp160 / chemistry
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HIV Envelope Protein gp160 / genetics
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HIV-1 / drug effects*
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HIV-1 / genetics*
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HIV-1 / pathogenicity
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HIV-1 / physiology
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Humans
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Piperazines / pharmacology*
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Protein Structure, Tertiary
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Pyrimidines / pharmacology*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
Substances
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Anti-HIV Agents
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CCR5 Receptor Antagonists
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DNA Primers
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DNA, Viral
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HIV Envelope Protein gp120
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HIV Envelope Protein gp160
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Piperazines
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Pyrimidines
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Recombinant Fusion Proteins
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gp120 protein, Human immunodeficiency virus 1
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gp160 protein, Human immunodeficiency virus 1
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vicriviroc
Associated data
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GENBANK/EU090200
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GENBANK/EU090201
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GENBANK/EU090202
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GENBANK/EU090203
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GENBANK/EU090204
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GENBANK/EU090205
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GENBANK/EU090206
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GENBANK/EU090207
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GENBANK/EU090208
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GENBANK/EU090209
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GENBANK/EU090210
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GENBANK/EU090211
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GENBANK/EU090212
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GENBANK/EU090213
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GENBANK/EU090214
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GENBANK/EU090215
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GENBANK/EU090216
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GENBANK/EU090217
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GENBANK/EU090218
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GENBANK/EU090219
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GENBANK/EU090220
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GENBANK/EU090221
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GENBANK/EU090222
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GENBANK/EU090223