EBV-induced gene 3 transcription is induced by TLR signaling in primary dendritic cells via NF-kappa B activation.
The EBV-induced gene 3 (EBI3) is expressed in dendritic cells (DCs) and part of the cytokine IL-27 that controls Th cell development. However, its regulated expression in DCs is poorly understood. In the present study we demonstrate that EBI3 is expressed in splenic CD8(-), CD8(+), and plasmacytoid DC subsets and is induced upon TLR signaling. Cloning and functional analysis of the EBI3 promoter using in vivo footprinting and mutagenesis showed that stimulation via TLR2, TLR4, and TLR9 transactivated the promoter in primary DCs via NF-kappaB and Ets binding sites at -90 and -73 bp upstream of the transcriptional start site, respectively. Furthermore, we observed that NF-kappaB p50/p65 and PU.1 were sufficient to transactivate the EBI3 promoter in EBI3-deficient 293 cells. Finally, induced EBI3 gene expression in DCs was reduced or abrogated in TLR-2/TLR4, TLR9, and MyD88 knockout mice, whereas both basal and inducible EBI3 mRNA levels in DCs were strongly suppressed in NF-kappaB p50-deficient mice. In summary, these data suggest that EBI3 expression in DCs is transcriptionally regulated by TLR signaling via MyD88 and NF-kappaB. Thus, EBI3 gene transcription in DCs is induced rapidly by TLR signaling during innate immune responses preceding cytokine driven Th cell development.
Wirtz
Stefan
S
Laboratory of Immunology, First Medical Clinic, University of Mainz, Mainz, Germany.
Becker
Christoph
C
Fantini
Massimo C
MC
Nieuwenhuis
Edward E
EE
Tubbe
Ingrid
I
Galle
Peter R
PR
Schild
Hans-Jörg
HJ
Birkenbach
Mark
M
Blumberg
Richard S
RS
Neurath
Markus F
MF
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008562:Membrane Glycoproteins / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism* / Q000502:physiology; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D021122:Protein Subunits / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D012315:RNA Caps / Q000235:genetics; D011956:Receptors, Cell Surface / Q000502:physiology*; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D015854:Up-Regulation / Q000276:immunology
C499601:Ebi3 protein, mouse; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D015778:Minor Histocompatibility Antigens; D016328:NF-kappa B; D021122:Protein Subunits; D012315:RNA Caps; D011956:Receptors, Cell Surface; D018121:Receptors, Cytokine; C495342:TLR2 protein, human; C495345:TLR4 protein, human; C495371:TLR9 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors