Induction of human dendritic cell maturation using transfection with RNA encoding a dominant positive toll-like receptor 4

Robin M. Cisco, Zeinab Abdel-Wahab, Jens Dannull, Smita Nair, Douglas Tyler, Eli Gilboa, Johannes Vieweg, Yehia Daaka, Scott K. Pruitt

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53 Scopus citations

Abstract

Maturation of dendritic cells (DC) is critical for the induction of Ag-specific immunity. Ag-loaded DC matured with LPS, which mediates its effects by binding to Toll-like receptor 4 (TLR4), induce Ag-specific CTL in vitro and in vivo in animal models. However, clinical use of LPS is limited due to potential toxicity. Therefore, we sought to mimic the maturation-inducing effects of LPS on DC by stimulating TLR4-mediated signaling in the absence of exogenous LPS. We developed a constitutively active TLR4 (caTLR4) and demonstrated that transfection of human DC with RNA encoding caTLR4 led to IL-12 and TNF-α secretion. Transfection with caTLR4 RNA also induced a mature DC phenotype. Functionally, transfection of DC with caTLR4 RNA enhanced allostimulation of CD4+ T cells. DC transfected with RNA encoding the MART (Melan-A/MART-1) melanoma Ag were then used to stimulate T cells in vitro. Cotransfection of these DC with caTLR4 RNA enhanced the generation of MART-specific CTL. This CTL activity was superior to that seen when DC maturation was induced using either LPS or a standard mixture of cytokines (TNF-α, IL-6, IL-1β, and PGE2). We conclude that transfection of DC with RNA encoding a functional signaling protein, such as caTLR4, may provide a new tool for studying TLR signaling in DC and may be a promising approach for the induction of DC maturation for tumor immunotherapy.

Original languageEnglish (US)
Pages (from-to)7162-7168
Number of pages7
JournalJournal of Immunology
Volume172
Issue number11
StatePublished - Jun 1 2004
Externally publishedYes

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ASJC Scopus subject areas

  • Immunology

Cite this

Cisco, R. M., Abdel-Wahab, Z., Dannull, J., Nair, S., Tyler, D., Gilboa, E., Vieweg, J., Daaka, Y., & Pruitt, S. K. (2004). Induction of human dendritic cell maturation using transfection with RNA encoding a dominant positive toll-like receptor 4. Journal of Immunology, 172(11), 7162-7168.