Characterization of a human immunodeficiency virus type 1 V3 deletion mutation that confers resistance to CCR5 inhibitors and the ability to use aplaviroc-bound receptor.
The human immunodeficiency virus type 1 (HIV-1) V3 loop is essential for coreceptor binding and principally determines tropism for the CCR5 and CXCR4 coreceptors. Using the dual-tropic virus HIV-1(R3A), we previously made an extensive panel of V3 deletions and identified subdomains within V3 that could differentially mediate R5 and X4 tropism. A deletion of residues 9 to 12 on the N-terminal side of the V3 stem ablated X4 tropism while leaving R5 tropism intact. This mutation also resulted in complete resistance to several small-molecule CCR5 inhibitors. Here, we extend these studies to further characterize a variant of this mutant, Delta9-12a, adapted for growth in CCR5(+) SupT1 cells. Studies using coreceptor chimeras, monoclonal antibodies directed against the CCR5 amino terminus (NT) and extracellular loops, and CCR5 point mutants revealed that, relative to parental R3A, R5-tropic Delta9-12a was more dependent on the CCR5 NT, a region that contacts the gp120 bridging sheet and V3 base. Neutralization sensitivity assays showed that, compared to parental R3A, Delta9-12a was more sensitive to monoclonal antibodies b12, 4E10, and 2G12. Finally, cross-antagonism assays showed that Delta9-12a could use aplaviroc-bound CCR5 for entry. These studies indicate that increased dependence on the CCR5 NT represents a mechanism by which HIV envelopes acquire resistance to CCR5 antagonists and may have more general implications for mechanisms of drug resistance that arise in vivo. In addition, envelopes such as Delta9-12a may be useful for developing new entry inhibitors that target the interaction of gp120 and the CCR5 NT.
Nolan
Katrina M
KM
Department of Medicine, Hematology-Oncology Division, University of Pennsylvania, Philadelphia, PA 19104, USA.
Del Prete
Gregory Q
GQ
Jordan
Andrea P O
AP
Haggarty
Beth
B
Romano
Josephine
J
Leslie
George J
GJ
Hoxie
James A
JA
D000911:Antibodies, Monoclonal / Q000276:immunology; D001565:Benzoates / Q000494:pharmacology*; D002460:Cell Line; D054659:Diketopiperazines; D024882:Drug Resistance, Viral; D015483:HIV Antibodies / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000235:genetics* / Q000378:metabolism; D023581:HIV Fusion Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010879:Piperazines / Q000494:pharmacology*; D011485:Protein Binding; D019713:Receptors, CCR5 / Q000502:physiology; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D013141:Spiro Compounds / Q000494:pharmacology*; D053586:Virus Internalization / Q000187:drug effects*
D000911:Antibodies, Monoclonal; D001565:Benzoates; D054659:Diketopiperazines; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D023581:HIV Fusion Inhibitors; D010879:Piperazines; D019713:Receptors, CCR5; D013141:Spiro Compounds; C514793:gp120 protein, Human immunodeficiency virus 1; C499671:aplaviroc