I tried 48 h pre-incubation with rec. m. IFN gamma (should increase IL-31RA at least in human monocyte derived DC). Then stimulation with rec. m. IL-31, but I could not detect any TNF as described for human DC. LPS leads to expected TNF "explosion".
Any ideas? My next step would be IL-6 + low (low!!) dose of LPS.
Thanks
Wolfgang
Hi Wolfgang,
Have you considered STAT phosphorylation as read out? We are doing lots of cytokine-induced phospho-flow assays in our lab, albeit using primary human T cells mostly. We haven't tried out IFNg but type 1 IFNs result in robust STAT1 phosphorylation within minutes and is easily detectable by flow cytometry. Similarly, IL-6 induces pSTAT3 very potently in T cells (and monocytes). However, I would not think that DCs respond well to 'normal' (classical) IL-6 signaling. Although activated DCs produce a lot of IL-6, they do not express (much) of the IL-6 receptor (IL-6R) on their surface. They are, however, susceptible to IL-6 'transsignaling' using a complex of IL-6 with soluble IL-6R...still, to test your JAK inhibitors, I would probably go for type I INF (via JAK1/TYK2) or IFNg (via JAK1/JAK2) induced pSTAT1.
Hope this is useful.
Christian
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology