Designing and engineering of DNA-vaccine construction encoding multiple CTL-epitopes of major HIV-1 antigens

Vaccine. 2004 Apr 16;22(13-14):1672-82. doi: 10.1016/j.vaccine.2003.09.048.

Abstract

A synthetic T cell immunogen (TCI) has been designed as a candidate DNA-based vaccine against Human immunodeficiency virus (HIV)-1 using cytotoxic T lymphocytes (CD8(+) CTL) and T-helper lymphocytes (CD4(+) Th) epitopes retrieved from the Los Alamos HIV Molecular Immunology Database. The protein 392 amino acids in length contains about eighty CTL-epitopes, many of which are overlapping and are totally restricted by ten different HLA class I molecules. To be able to detect CTL responses induced by a DNA vaccine in experimental animals, additional epitopes, restricted by mouse and Macaque rhesus major histocompatibility complex (MHC) class I molecules, were included in the target immunogen. The gene encoding the TCI protein was assembled, cloned into vector plasmids and expressed in a prokaryotic and a eukaryotic system. The presence of HIV-1 protein fragments in the immunogen structure was ascertained by ELISA and immunoblotting using panels of HIV-1-positive sera and monoclonal antibodies to p24. It has been demonstrated that DNA vaccine can induce both specific T cell responses (CTL and blast transformation) and specific antibodies in mice immunized with pcDNA-TCI.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / genetics*
  • AIDS Vaccines / immunology*
  • Animals
  • Antibody Specificity
  • Base Sequence
  • Cell Division
  • DNA, Viral / genetics
  • DNA, Viral / immunology
  • Drug Design
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / genetics*
  • Epitopes / immunology*
  • Escherichia coli / genetics
  • Genetic Engineering
  • HIV Antibodies / analysis
  • HIV Antibodies / biosynthesis
  • HIV Antigens / genetics*
  • HIV Antigens / immunology*
  • HIV-1 / genetics*
  • HIV-1 / immunology*
  • Humans
  • Immunochemistry
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Plasmids / immunology
  • T-Lymphocytes, Cytotoxic / immunology*
  • Vaccines, DNA / immunology

Substances

  • AIDS Vaccines
  • DNA, Viral
  • Epitopes
  • HIV Antibodies
  • HIV Antigens
  • Vaccines, DNA