RNA interference shows critical requirement for NF-kappa B p50 in the production of IL-12 by human dendritic cells.
Specific NF-kappaB/Rel proteins regulate murine dendritic cell (DC) survival, differentiation, and activation, but little is known of their role in human cells because of limited loss-of-function analyses. RNA interference (RNAi) is a mechanism to effectively silence gene expression via sequence-specific double-stranded small interfering RNAs (siRNAs). RNAi was used to assess the role of the p50 (NF-kappaB1) protein in the maturation and activation of cultured human monocyte-derived DC (MoDC). Transfection of cultured MoDC with siRNAs reduced p50 mRNA and protein levels in a specific, dose-dependent, and time-dependent manner. Basal or maturation-induced expression of HLA-DR and costimulatory molecules were not affected, whereas transcription of the IL-12 p40 gene and the secretion of IL-12alphabeta were reduced. Such MoDC induced less IFN-gamma production by T cells in MLR. This is the first report of RNAi-induced phenotype in human primary DC with a method that caused no measurable toxicity or type-I IFN response. siRNAs appear useful for the study of signaling pathways in immune cells, revealing a pivotal requirement for p50 in MoDC for IL-12 production and induction of optimal type-1 immune responses.
Laderach
Diego
D
Institut National de la Santé et de la Recherche Médicale Unité 362, Institut Gustave Roussy, Villejuif, France.
Compagno
Daniel
D
Danos
Olivier
O
Vainchenker
William
W
Galy
Anne
A
D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D015536:Down-Regulation / Q000235:genetics / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007369:Interferon Inducers / Q000633:toxicity; D007370:Interferon Type I / Q000096:biosynthesis; D018664:Interleukin-12 / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D052002:NF-kappa B p50 Subunit; D034622:RNA Interference / Q000276:immunology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D034741:RNA, Small Interfering / Q000494:pharmacology* / Q000633:toxicity; D018417:Th1 Cells / Q000276:immunology / Q000378:metabolism
D007369:Interferon Inducers; D007370:Interferon Type I; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018664:Interleukin-12