I have been using THP1 cell lines. We use PMA (24 h treatment) for the differentiation of Monocytes into macrophages. But in my experiments, when I stimulate the macrophages with cytokines, i do not get the required results. The results are aberrant, like the changes in cytokine production are not in an order. After 24 hr PMA treatment, in our lab, we do not change the media (having PMA) and directly add the specific cytokine for investigating their effects.
Here, i want to know if the PMA is media is interefering with the stimultatory effect of our cytokine? I couldn't find any literature. If there is any literature evidence, please share.
PMA is a strong stimulator of lymphocytes and monocytes and induces release of a variety of cytokines. it may minimize the effects of the cytokines you add.
Thank you so much for your answer. Is there any literature, supporting this statement? I had also assumed this but didn't find any literature for this.
PMA along with brefeldin A (to inhibit complete release) is widely used in flow cytometry studies to detect intracelular cytokines because of this effect. I am talking about normal monocytes though, not in vitro differentiated macrophages
Phorbol esters end up activating JNKs and subsequently AP-1 that controls expression of a lot of cytokines
THP-1 differentiated macrophages can be further activated through TLRs ( i.e. LPS) but I do not know about cytokines
If I found a suitable reference I will share it with you
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