Induction of human immunodeficiency virus neutralizing antibodies using fusion complexes

Donato Zipeto, Andrea Matucci, Chiara Ripamonti, Gabriella Scarlatti, Paola Rossolillo, Marco Turci, Silvia Sartoris, Giuseppe Tridente, Umberto Bertazzoni

Research output: Contribution to journalArticlepeer-review

Abstract

Human immunodeficiency virus-1 (HIV-1) infects cells by membrane fusion that is mediated by the envelope proteins gp120/gp41 and the cellular receptors CD4 and CCR5. During this process, some conserved viral epitopes are temporarily exposed and may induce a neutralizing antibody response when fixed in the fusogenic conformation. These transient structures are conserved and may be effective antigens for use in an anti-HIV-1 vaccine. In this study we tested different conditions of preparation of fusion complexes inducing neutralizing antibodies against both R5 and X4 tropic HIV-1 strains. Cell lines expressing HIV-1 gp120/gp41 and CD4-CCR5 were prepared and conditions for producing fusion complexes were tested. Complexes produced at different temperature and fixative combinations were used to immunize mice. Results indicated that (a) fusion complexes prepared at either 21 °C, 30 °C or 37 °C were immunogenic and induced neutralizing antibodies against both R5 and X4 HIV-1 heterologous isolates; (b) after extensive purification of antibodies there was no cytotoxic effect; (c) complexes prepared at 37 °C were more immunogenic and induced higher titers of neutralizing antibodies than complexes prepared at either 21 °C or 30 °C; (d) the fixative used did not affect the titer of neutralizing antibodies except for glutaraldehyde which was ineffective; (e) the neutralizing activity was retained after CD4-CCR5 antibody removal. The production of higher titers of neutralizing antibody with fusion complexes prepared at 37 °C, as compared to lower temperatures, may be related to the induction of antibodies against many different conformation intermediates that subsequently act synergistically at different steps in the fusion process.

Original languageEnglish
Pages (from-to)1424-1433
Number of pages10
JournalMicrobes and Infection
Volume8
Issue number6
DOIs
Publication statusPublished - May 2006

Keywords

  • Fusion complex
  • Membrane fusion
  • Neutralizing antibodies

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Induction of human immunodeficiency virus neutralizing antibodies using fusion complexes'. Together they form a unique fingerprint.

Cite this