Human immunodeficiency virus type 1 entry inhibitors selected on living cells from a library of phage chemokines.
The chemokine receptors CCR5 and CXCR4 are promising non-virus-encoded targets for human immunodeficiency virus (HIV) therapy. We describe a selection procedure to isolate mutant forms of RANTES (CCL5) with antiviral activity considerably in excess of that of the native chemokine. The phage-displayed library of randomly mutated and N-terminally extended variants was screened by using live CCR5-expressing cells, and two of the selected mutants, P1 and P2, were further characterized. Both were significantly more potent HIV inhibitors than RANTES, with P2 being the most active (50% inhibitory concentration of 600 pM in a viral coat-mediated cell fusion assay, complete protection of target cells against primary HIV type 1 strains at a concentration of 10 nM). P2 resembles AOP-RANTES in that it is a superagonist of CCR5 and potently induces receptor sequestration. P1, while less potent than P2, has the advantage of significantly reduced signaling activity via CCR5 (30% of that of RANTES). Additionally, both P1 and P2 exhibit not only significantly increased affinity for CCR5 but also enhanced receptor selectivity, retaining only trace levels of signaling activity via CCR1 and CCR3. The phage chemokine approach that was successfully applied here could be adapted to other chemokine-chemokine receptor systems and used to further improve the first-generation mutants reported in this paper.
Hartley
Oliver
O
Département de Biochimie Médicale, Centre Médical Universitaire, Geneva, Switzerland.
Dorgham
Karim
K
Perez-Bercoff
Danielle
D
Cerini
Fabrice
F
Heimann
Anouk
A
Gaertner
Hubert
H
Offord
Robin E
RE
Pancino
Gianfranco
G
Debré
Patrice
P
Gorochov
Guy
G
D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D001667:Binding, Competitive; D016466:CHO Cells; D002460:Cell Line; D018946:Chemokine CCL5 / Q000031:analogs & derivatives* / Q000235:genetics / Q000494:pharmacology*; D018925:Chemokines / Q000235:genetics / Q000494:pharmacology*; D006224:Cricetinae; D015497:HIV-1 / Q000187:drug effects / Q000472:pathogenicity*; D006801:Humans; D008561:Membrane Fusion / Q000187:drug effects; D009154:Mutation; D019151:Peptide Library; D019713:Receptors, CCR5 / Q000378:metabolism
D019380:Anti-HIV Agents; D018946:Chemokine CCL5; D018925:Chemokines; D019151:Peptide Library; D019713:Receptors, CCR5; C111989:aminooxypentane-RANTES