Enhanced dendritic cell antigen presentation in RNA-based immunotherapy

Matthew F. Kalady, Mark W. Onaitis, Karen M. Padilla, Sirisha Emani, Douglas S. Tyler, Scott K. Pruitt

Research output: Contribution to journalArticlepeer-review

55 Scopus citations


Background. Dendritic cells pulsed with mRNA provide a unique approach to tumor immunotherapy. We hypothesized that increased mRNA transfection efficiency and dendritic cell maturation would improve antigen processing and presentation as well as T-cell costimulation, resulting in enhanced induction of antimelanoma immune responses. Methods. Immature monocyte-derived dendritic cells were transfected with mRNA by passive pulsing, lipofection, or electroporation. Dendritic cells were either left untreated or matured using the double-stranded RNA poly(I:C). T-Cell cultures were generated by stimulation of naïve T-cells with each set of dendritic cells. Specific antigen presentation and specific effector T-cell generation were analyzed by an IFN-γ release Elispot assay. Results. Greatest intracellular green fluorescent protein was observed by flow cytometry following dendritic cell electroporation with green fluorescent protein mRNA. DC presentation of Mart-1/Melan A peptide, as measured by Elispot assay using a specific T-cell clone, was greatest following transfection with Mart-1/Melan A mRNA by electroporation. Maturation of dendritic cells further improved antigen presentation regardless of transfection technique. Specific Mart-1/Melan A effector T cells were produced after culture of naïve T cells with dendritic cells that were electroporated with Mart-1/Melan A mRNA and then matured, but not for dendritic cells that remained immature. Conclusions. Efficient mRNA transfection by electroporation as well as dendritic cell maturation results in increased levels of Mart-1/Melan A antigen presentation and enhanced production of antigen-specific effector T cells. This combination of strategies may be used to enhance immune responses to RNA-based dendritic cell vaccines.

Original languageEnglish (US)
Pages (from-to)17-24
Number of pages8
JournalJournal of Surgical Research
Issue number1
StatePublished - 2002
Externally publishedYes


  • Antigen presentation
  • Dendritic cells
  • Immunotherapy
  • mRNA-based vaccines

ASJC Scopus subject areas

  • Surgery


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