Characterization of immune responses induced by combined clade-A HIV-1 recombinant adenovectors in mice

Iran J Immunol. 2010 Sep;7(3):162-76.

Abstract

Background: Numerous evidences indicate that in some HIV-1 positive patients, the humoral and cellular immune responses are induced against HIV-1 proteins and this is inversely related to the progress of infection.

Objective: The aim of this study was the evaluation of the Adenovectors containing HIV genes in induction of immune responses in mice.

Methods: The HIV-1 genes including gag p24, rev, nef and exon-1 of tat were amplified from HIV-1 RNA (clade-A). The cDNA of each gene was cloned into a transfer vector. The transfer vector was then co-transformed into E. coli strain BJ5183 together with pAdenovector ∆E1/E3. The recombinant adenoviral construct was transfected into QBI-293A cells. Recombinant viruses were purified and titrated on 293 cell plates. Expression of transgenes was evaluated using western blotting. Then 10(12) viral particles were injected into 15 groups of 5 mice and all patterns of combination of these 4 HIV-1 genes were evaluated. After 2 weeks, humoral and cellular immune responses were evaluated using ELISA, cell proliferation and ELISpot (IL-2, IL-4 and IFN-γ) assays, consecutively.

Results: It was demonstrated that each gene was expressed. The response targets were mostly toward Th1, though several Th2 responses were also observed. Single injection in our study induced a good cellular response but the humoral responses were not as strong as the cellular ones.

Conclusion: Considering and comparing all results and evaluating the various possible interactions revealed that simultaneous injection of tat and gag has enhanced the humoral and cellular responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / genetics
  • AIDS Vaccines / immunology
  • Adenoviridae / genetics*
  • Adenoviridae / immunology*
  • Animals
  • Cytokines / metabolism
  • Female
  • Gene Expression
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics*
  • Genetic Vectors / immunology*
  • Genotype*
  • HIV Antibodies / blood
  • HIV Antibodies / immunology
  • HIV Antigens / genetics
  • HIV Antigens / immunology
  • HIV Infections / immunology
  • HIV Infections / metabolism
  • HIV Infections / prevention & control
  • HIV-1 / genetics*
  • HIV-1 / immunology*
  • Humans
  • Immunity, Cellular
  • Immunity, Humoral
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Lymphocyte Activation / immunology
  • Mice
  • Spleen / immunology
  • Spleen / metabolism
  • Transgenes
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology
  • Viral Proteins / genetics
  • Viral Proteins / immunology

Substances

  • AIDS Vaccines
  • Cytokines
  • HIV Antibodies
  • HIV Antigens
  • Immunoglobulin G
  • Vaccines, DNA
  • Viral Proteins