Membrane-Anchored Peptide Inhibits Human Immunodeficiency Virus Entry

Abstract

Peptides derived from the heptad repeats of human immunodeficiency virus (HIV) gp41 envelope glycoprotein, such as T20, can efficiently inhibit HIV type 1 (HIV-1) entry. In this study, replication of HIV-1 was inhibited more than 100-fold in a T-helper cell line transduced with a retrovirus vector expressing membrane-anchored T20 on the cell surface. Inhibition was independent of coreceptor usage.

C-terminal Octylation Rescues an Inactive T20 Mutant

“…(i) T20 lacks the residues that bind to the coiled coil C-terminal cavity; (ii) the known crystal and solution structures of fragments of HIV or SIV gp41 ectodomains do not fully include the regions corresponding either to T20 or to its putative binding site at the N terminus of the coiled coil; and (iii) in addition to a primary target site within the N-terminal heptad repeat (21), T20 has been postulated to bind to a second site in gp41, presumably a non-specified region at the C terminus of the ectodomain close to the viral membrane (22,23). Indeed, that T20 acts in close proximity to the membrane is supported by the studies of von Laer and co-workers (24). They have shown that when T20 is anchored to the target cell membrane by fusion to a transmembrane domain, the wild type and the ANAA mutant T20 have similar inhibitory activities.…”

Gene therapy progress and prospects: Novel gene therapy approaches for AIDS

“…The T20 peptide, when expressed on the surface of PM-1 cells as an anchored-membrane construct, effectively inhibited HIV-1 entry. 53 A retroviral vector, the peptide itself, and a scaffold for its presentation were optimized for maximal expression and localization on the cell surface. The membrane-anchored C-peptide binds to the free version of gp41 N peptides, suggesting that the membraneanchored C peptides exert their biological effect by binding gp41.…”

Efficient entry inhibition of human and nonhuman primate immunodeficiency virus by cell surface-expressed gp41-derived peptides

“…8,9 M87o-RRE vector was engineered to contain the HIV C46 sequence (M87o HIV ). Generation of the homologous M87o SIV and M87o SHIV as well as the low-expressing vectors M87o HIV low, M87o SIV low and M87o SHIV low is described in detail in the Supplementary materials and methods.…”

“…47 Low MOI were used for transduction of PM-1 cells to obviate multiple vector integrations. Cells stained with the human mAb 2F5 (kindly provided by H Katinger) directed against a motif in the C-peptide of gp41 8 and a monoclonal goat anti-human immunoglobulin G antibody coupled to phycoerythrin (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) were analyzed by flow cytometry to assess transduction efficiency. PM-1 cells, transduced with the vectors that contain a neomycin resistance gene, were selected with G418 (0.8 mg ml À1 ) for 10 days.…”

“…8 Optimization of the retroviral vector, the scaffold for presentation of the peptide on the cell surface and elongation of the C36-to a C46-peptide (M87o) have maximized expression of maC46 on PM-1 cells and primary lymphocytes. In addition antiretroviral activity against a broad range of viral isolates was enhanced.…”