Dendritic cells transfected with interleukin-12 and tumor-associated antigen messenger RNA induce high avidity cytotoxic T cells

Gene Ther. 2007 Feb;14(4):366-75. doi: 10.1038/sj.gt.3302874. Epub 2006 Oct 12.

Abstract

Dendritic cells (DC) transfected with messenger RNA (mRNA) encoding tumor-associated antigens (TAA) are able to induce potent tumor-specific T-cell responses directed to a broad spectrum of tumor-associated epitopes. The in vitro generation of DC possessing all the features crucial for the induction of type 1 immune responses, such as mature state, migratory potential and interleukin-12 (IL-12p70) production is complicated. Particularly migratory potential is inversely correlated with IL-12p70 production after maturation with prostaglandin E2 (PGE2), which is included in maturation cocktails currently used in most vaccination trials. Here, we show that transfection of PGE2 matured DC with a single mRNA strain encoding for ubiquitin followed by a TAA which was linked to IL-12 by a self-cleaving 2A sequence, produced biological active IL-12p70 and were able to present the transfected TAA up to 72 h after transfection. Furthermore, use of the anti-reverse cap analog for in vitro transcription of the IL-12 mRNA enabled constitutive IL-12p70 production for up to 5 days. These transfected mature DC migrated efficiently towards lymph node derived chemokines. DCs constitutively expressing IL-12p70, generate TAA-specific cytotoxic T cells with an high functional avidity, independent of CD4+ T-cell help.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / genetics*
  • Cancer Vaccines
  • Cells, Cultured
  • Cytotoxicity Tests, Immunologic
  • Cytotoxicity, Immunologic
  • Dendritic Cells / immunology*
  • Epitopes / immunology
  • Genetic Therapy / methods*
  • Humans
  • Immunophenotyping
  • Immunotherapy, Adoptive / methods*
  • Interleukin-12 / genetics*
  • Interleukin-12 / immunology
  • Lymphocyte Activation
  • MART-1 Antigen
  • Neoplasm Proteins / genetics*
  • RNA, Messenger / administration & dosage
  • RNA, Messenger / genetics*
  • T-Lymphocytes, Cytotoxic / immunology
  • Transfection / methods

Substances

  • Antigens, Neoplasm
  • Cancer Vaccines
  • Epitopes
  • MART-1 Antigen
  • MLANA protein, human
  • Neoplasm Proteins
  • RNA, Messenger
  • Interleukin-12