Induction of human dendritic cell maturation using transfection with RNA encoding a dominant positive toll-like receptor 4.
ISBN or ISSN
Journal of immunology
Publication Date / Copyright Date
American Association of Immunologists
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-alpha 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-alpha, IL-6, IL-1beta, and PGE(2)). 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.
Medicine and Health Sciences
Cell Line, Dendritic Cells, Granulocytes, Humans, Interleukin-12, Isoantigens, Lipopolysaccharides, Membrane Glycoproteins, RNA, Receptors, Cell Surface, Signal Transduction, T-Lymphocytes, Toll-Like Receptor 4, Toll-Like Receptors, Transfection, Tumor Necrosis Factor-alpha
Cisco, Robin M; Abdel-Wahab, Zeinab; Dannull, Jens; Nair, Smita; Tyler, Douglas S; Gilboa, Eli; Vieweg, Johannes; Daaka, Yehia; and Pruitt, Scott K, "Induction of human dendritic cell maturation using transfection with RNA encoding a dominant positive toll-like receptor 4." (2004). NSU-MD Faculty Articles. 32.