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| Status |
Public on Nov 13, 2013 |
| Title |
Real-time quantitative PCR analysis reveals high expression of activin A during NK-DC interactions |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
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| Summary |
The interaction of natural killer (NK) cells with dendritic cells (DC) results in reciprocal cell activation through the interaction of membrane proteins and the release of soluble factors. Here we report that in NK-DC cocultures, among a set of 84 cytokines investigated, activin A was the second highest induced gene, with CXCL8 being the most upregulated one. Activin A is a member of the TGF-β superfamily and was previously shown to possess both pro- and antiinflammatory activities. In NK-DC cocultures, the induction of activin A required cell contact and was dependent on the presence of proinflammatory cytokines (i.e. IFN-γ, TNF-α and GM-CSF) as well as on NK cell-mediated DC killing. CD1+ DC were the main activin A producer cells among myeloid blood DC subsets. In NK-DC cocultures, inhibition of acitivn A by follistatin, a natural inhibitory protein, or by a specific blocking antibody, resulted in the upregulation of proinflammatory cytokine release (i.e. IL-6, IL-8, TNF-α) by DC and in the increase of DC maturation. In conclusion, our study reports that activin A, produced during NK-DC interactions, represents a relevant negative feedback mechanism that might function to prevent excessive immune activation by DC.
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| Overall design |
Human CD14 positive monocytes were differentiated to DC in vitro in the presence of IL-4 (20 ng/ml) and GM-CSF (50 ng/ml) for 6 days. Immature DC were then cocultured with allogeneic IL-15-activated NK cells (at 1:1 NK:DC ratio) for 0, 2 and 6 hrs. RNA was obtained from three independent coculture experiments. Equal amount of total RNA from each experiment was pooled prior to gene expression analysis. The gene expression of common cytokines was quantified using an RT2 Profiler PCR Array (Qiagen).
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| Contributor(s) |
Seeger P, Bosisio D, Parolini S, Badolato R, Gismondi A, Santoni A, Sozzani S |
| Citation(s) |
24395917 |
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| Submission date |
Nov 12, 2013 |
| Last update date |
Feb 16, 2014 |
| Contact name |
Pascal Seeger |
| Organization name |
University of Brescia
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| Department |
Department of Molecular and Translational Medicine
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| Street address |
Viale Europa 11
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| City |
Brescia |
| ZIP/Postal code |
25123 |
| Country |
Italy |
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| Platforms (1) |
| GPL17916 |
Human Common Cytokines RT² Profiler™ PCR Array |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA227367 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE52306_fold-change.txt.gz |
728 b |
(ftp)(http) |
TXT |
| GSE52306_non-normalized.txt.gz |
658 b |
(ftp)(http) |
TXT |
| Processed data provided as supplementary file |
| Processed data are available on Series record |
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