Identification of HIV vaccine candidate peptides by screening random phage epitope libraries

Virology. 1993 Apr;193(2):709-16. doi: 10.1006/viro.1993.1179.

Abstract

Most synthetic HIV-1 gp120 V3 loop peptides that are used as immunogens in experimental HIV-1 vaccine studies are modeled from the naturally occurring viral gp120 V3 loops. In experimental animals these immunogens generally elicit type (or variant)-specific neutralizing antibodies that are not broadly reactive among HIV-1 variants. In an attempt to find a more general structure for the V3 loop, we have obtained candidates that mimic V3 loop sequences by screening random epitopes displayed in a fusion phage 15-residue epitope library. Human monoclonal antibody 447-52D, a highly potent and broadly reactive virus-neutralizing antibody that recognizes the conserved V3 loop tip motif GPXR, was the probe. By using a screening method that was designed specifically for this work, we identified hundreds of reactive phage clones, 70 of which were sequenced. Over 98% of the epitopes contain the motif GPXR, yet none of the 70 are an identical match to any V3 variant loop described to date. One of these sequences was synthesized as the beta-maleimidopropionyl 15-mer peptide, covalently conjugated to a carrier and used to immunize rabbits. High anti-peptide titers were obtained in all animals with three of four individual responses also binding to a peptide that is representative of the "North American consensus" V3 loop. The sera from these three positive rabbits neutralized HIV-1 variant SF-2 in vitro. In addition, one of them was capable of neutralizing variant AL-1. Both of these variants are considered to have V3 loops of the North American consensus type. Thus, neutralizing responses were obtained by use of an immunogen that was selected for its ability to bind a broadly reactive human monoclonal antibody rather than modeled from an HIV-1 gp120 V3 loop sequence.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • AIDS Vaccines*
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Base Sequence
  • Epitopes / immunology*
  • Escherichia coli / genetics
  • Genetic Variation
  • HIV Envelope Protein gp120 / immunology*
  • HIV-1 / genetics
  • HIV-1 / immunology*
  • Humans
  • Molecular Sequence Data
  • Neutralization Tests
  • Oligodeoxyribonucleotides
  • Peptides / chemical synthesis
  • Polymerase Chain Reaction / methods
  • Rabbits / immunology
  • Vaccines, Synthetic*

Substances

  • AIDS Vaccines
  • Antibodies, Monoclonal
  • Epitopes
  • HIV Envelope Protein gp120
  • Oligodeoxyribonucleotides
  • Peptides
  • Vaccines, Synthetic