Coordinate NF-kappaB and STAT1 activation promotes development of myeloid type 1 dendritic cells.
NF-kappaB and STAT1 are critically involved in the initiation of the inflammatory cascade. Using semi-automated imaging cytometry and fluorescent antibodies, we screened several factors for their ability to induce nuclear translocation of RelA/NF-kappaB and STAT1 in subsets of monocyte-derived dendritic cells (DC). Detailed kinetics and dose-response studies are presented for IL-1-, LPS-, CD40L-, IFN-gamma- and IFN-alpha-stimulated responses. The results are consistent with the notion that simultaneous activation of both STAT1 and NF-kappaB pathways at the initiation of differentiation culture is required for efficient priming of IL-12 production by DC. Maturation of DC led to characteristic NF-kappaB and STAT1 distribution and response patterns. During the resting stage, DC differentiated under the presence of IFN-gamma showed sustained STAT activation and remained responsive to LPS. By contrast, PGE2-supplemented DC could be characterized by negligible responses to LPS and IFN-gamma and a remarkable NF-kappaB response to CD40L. STAT1 pathway was suppressed in PGE2-supplemented cells. We conclude that the magnitude and temporal kinetics of the nuclear shift of STAT1 and NF-kappaB in myeloid DC are associated with IL-12p70 production and are dependent on the nature of the stimulating factors and the polarization state of cells.
Vakkila
J
J
Department of Surgery, G.27A Hillman Cancer Center, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA. jukka.vakkila@helsinki.fi
Demarco
R A
RA
Lotze
M T
MT
D002454:Cell Differentiation / Q000276:immunology*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D021621:Imaging, Three-Dimensional; D018664:Interleukin-12 / Q000096:biosynthesis; D022423:Myeloid Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D050794:STAT1 Transcription Factor / Q000378:metabolism*
D016328:NF-kappa B; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D018664:Interleukin-12