Synthetic multimeric peptides derived from the principal neutralization domain (V3 loop) of human immunodeficiency virus type 1 (HIV-1) gp120 bind to galactosylceramide and block HIV-1 infection in a human CD4-negative mucosal epithelial cell line.
The glycosphingolipid galactosylceramide (GalCer), which binds gp120 with high affinity and specificity, is a potential alternative receptor for human immunodeficiency virus type 1 (HIV-1) in some CD4-negative neural and epithelial human cells, including the human colonic epithelial cell line HT-29. In the present study, we demonstrate that synthetic multibranched peptides derived from the consensus sequence of the HIV-1 V3 loop block HIV-1 infection in HT-29 cells. The most active peptide was an eight-branched multimer of the motif Gly-Pro-Gly-Arg-Ala-Phe which at a concentration of 1.8 microM induced a 50% inhibition of HIV-1 infection in competition experiments. This peptide was not toxic to HT-29 cells, and preincubation with HIV-1 did not affect viral infectivity, indicating that the antiviral activity was not due to a nonspecific virucidal effect. Using a high-performance thin-layer chromatography binding assay, we found that multibranched V3 peptides recognized GalCer and inhibited binding of recombinant gp120 to the glycosphingolipid. In addition, these peptides abolished the binding of an anti-GalCer monoclonal antibody to GalCer on the surface of live HT-29 cells. These data provide additional evidence that the V3 loop is involved in the binding of gp120 to the GalCer receptor and show that multibranched V3 peptides are potent inhibitors of the GalCer-dependent pathway of HIV-1 infection in CD4-negative mucosal epithelial cells.
Yahi
N
N
CNRS URA 1455, Laboratoire de Biochimie, Faculté de Médecine Nord, Marseille, France.
Sabatier
J M
JM
Baghdiguian
S
S
Gonzalez-Scarano
F
F
Fantini
J
J
D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000378:metabolism*; D016384:Consensus Sequence; D005699:Galactosylceramides / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology / Q000821:virology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000276:immunology / Q000494:pharmacology*; D011485:Protein Binding; D014407:Tumor Cells, Cultured
D015704:CD4 Antigens; D005699:Galactosylceramides; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments