Hi Quyen,

In gerneral it depends what you want to observe.

To just show that your DC get activated it should be enough to measure CD80, CD86 and maybe CD40.

IL-6 is always produced in high abundancy. IL-10 on the other hand is only expressed in low amounts in mouse DC. You could also go for IL-1beta if you think there is something going on with the inflammasome.

Hope I could help.

Patrick

Could you quantify IL 12, which is a cytokine produced by antigen presenting cells and determines an effector Th1 response, inducing IFN-gamma producing T lymphocytes.

Hi Quyen,

your purpose is for stimulating CD8+ T cells or what?

you need to check the phenotypes for DC maturation, I think you should check: MHCI, MHCII, CD80, CD86, CD40, CCR7.

other hand for cytokines secretion, IL-12p70 and IL-10 that are enough.

plz follow these articals:

1, http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=5281&pubmed-linkout=1.

2, http://journals.lww.com/immunotherapy-journal/pages/articleviewer.aspx?year=2015&issue=10000&article=00004&type=abstract.

good luck! 

Dear,

Thank you very much for all your kindly answers.

I'm on the way to compare effect of different vaccine adjuvant on DC activation and maturation. And further study is on both CD4 and CD8 immune responses. I did check expression of CD40, 80, 86, MHC-II and measure IL12p70, IL1beta and TNF-alpha in vitro.

IL-10 is too low to detect when i treat DCs with adjuvants for 24h. Could you suggest incubation time for this.

I'm interested in DC migration also, i'm waiting to check expression of CCR7 first.

Thank you!

Maybe IL-10 is to low to detect, because there is no IL-10 induced? You should try to stimulate your DC with LPS or other potent activators, so that you have a positive control for IL-10 secretion.