Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef

Daniel G. Kavanagh, Daniel E. Kaufmann, Sherzana Sunderji, Nicole Frahm, Sylvie Le Gall, David Boczkowski, Eric S. Rosenberg, David R. Stone, Mary N. Johnston, Bradford S. Wagner, Mohammad T. Zaman, Christian Brander, Eli Gilboa, Bruce D. Walker, Nina Bhardwaj

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

Transfection with synthetic mRNA is a safe and efficient method of delivering antigens to dendritic cells for immunotherapy. Targeting antigens to the lysosome can sometimes enhance the CD4+ T-cell response. We transfected antigen-presenting cells (APCs) with mRNA encoding Gag-p24 and cytoplasmic, lysosomal, and secreted forms of Nef. Antigen-specific cytotoxic T cells were able to lyse the majority of transfected targets, indicating that transfection was efficient. Transfection of APCs with a Nef construct bearing lysosomal targeting signals produced rapid and prolonged antigen presentation to CD4+ and CD8+ T cells. Polyclonal CD4+ and CD8+ T-cell lines recognizing multiple distinct epitopes were expanded by coculture of transfected dendritic cells with peripheral blood mononuclear cells from viremic and aviremic HIV-infected subjects. Importantly, lysosome-targeted antigen drove a significantly greater expansion of Nef-specific CD4+ T cells than cytoplasmic antigen. The frequency of recognition of CD8 but not CD4 epitopes by mRNA-expanded T cells was inversely proportional to sequence entropy and was similar to ex vivo responses from a large chronic cohort. Thus human dendritic cells transfected with mRNA encoding lysosome-targeted HIV antigen can expand a broad, polyclonal repertoire of antiviral T cells, offering a promising approach to HIV immunotherapy.

Original languageEnglish (US)
Pages (from-to)1963-1969
Number of pages7
JournalBlood
Volume107
Issue number5
DOIs
StatePublished - Mar 1 2006
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Kavanagh, D. G., Kaufmann, D. E., Sunderji, S., Frahm, N., Le Gall, S., Boczkowski, D., Rosenberg, E. S., Stone, D. R., Johnston, M. N., Wagner, B. S., Zaman, M. T., Brander, C., Gilboa, E., Walker, B. D., & Bhardwaj, N. (2006). Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef. Blood, 107(5), 1963-1969. https://doi.org/10.1182/blood-2005-04-1513