Robust CD4+ and CD8+ T cell responses to SIV using mRNA-transfected DC expressing autologous viral Ag

Nada M. Melhem, Xiang Dong Liu, David Boczkowski, Eli Gilboa, Simon M. Barratt-Boyes

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

A potentially powerful strategy for theratic HIV vaccination is the use of DC transfected with mRNA encoding autologous viral Ag, as epitopes presented by transfected DC would exactly reflect those expressed by infected cells in the individual. Using human and rhesus macaque monocyte-derived DC, we show that nucleofection is a superior method for mRNA transfection, resulting in high-level protein expression and DC maturation. DC transfected with SIV gag isolated from an infected monkey stimulated robust Ag-specific recall T cell responses of similar magnitude to those induced by peptide-pulsed PBMC that were predominantly CD8+ T cell mediated. Enhanced CD4+ T cell responses were stimulated when Gag was redirected into the lysosomal pathway via the targeting signal derived from lysosome-associated membrane protein-1 (LAMP-1). Rhesus DC transfected with lysosome-targeted gag encoding an escape mutation in an immunodominant CTL epitope stimulated CD4+ and CD8+ T cell responses of almost equivalent magnitude directed towards undefined epitopes outside of the mutated region. Finally, gag-transfected DC from SIV-infected monkeys stimulated significant Ag-specific recall T cell responses in an entirely autologous system. These findings demonstrate that mRNA-transfected DC expressing SIV Ag derived from infected monkeys stimulate broad and relevant T cell responses, supporting this approach for therapeutic HIV vaccine development.

Original languageEnglish
Pages (from-to)2164-2173
Number of pages10
JournalEuropean Journal of Immunology
Volume37
Issue number8
DOIs
StatePublished - Aug 1 2007
Externally publishedYes

Fingerprint

T-Lymphocytes
Messenger RNA
Haplorhini
Epitopes
Lysosome-Associated Membrane Glycoproteins
Immunodominant Epitopes
AIDS Vaccines
Lysosomes
Macaca mulatta
Transfection
Monocytes
Vaccination
HIV
Peptides
Mutation
Proteins
Therapeutics

Keywords

  • DC
  • mRNA
  • SIV
  • T cell stimulation

ASJC Scopus subject areas

  • Immunology

Cite this

Robust CD4+ and CD8+ T cell responses to SIV using mRNA-transfected DC expressing autologous viral Ag. / Melhem, Nada M.; Liu, Xiang Dong; Boczkowski, David; Gilboa, Eli; Barratt-Boyes, Simon M.

In: European Journal of Immunology, Vol. 37, No. 8, 01.08.2007, p. 2164-2173.

Research output: Contribution to journalArticle

Melhem, Nada M. ; Liu, Xiang Dong ; Boczkowski, David ; Gilboa, Eli ; Barratt-Boyes, Simon M. / Robust CD4+ and CD8+ T cell responses to SIV using mRNA-transfected DC expressing autologous viral Ag. In: European Journal of Immunology. 2007 ; Vol. 37, No. 8. pp. 2164-2173.
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