I observe an increase in the PD-L1 expression of CD14+ cells after a special treatment. Now, I want to further analyze the PBMCs for changes in checkpoint protein expressions. I am designing a FACS panel, but I am not entirely sure what to include next. I thought about PD-L1, PD-1, CTLA-4, LAG-3, ICOS, CD3, CD4, CD14, and CD19. What do you think about including CD80 and CD86? Do you have any other suggestions?
Hi Katharina,
I assume that the CD14+ cells in your treatment were monocytes/macrophages. For this particular experiment, besides the markers you listed above, I would recommend including some of the cytokines that are usually associated with M1 and M2 macrophages.
You can find some of the information in this link: https://www.origene.com/research-areas/m1-and-m2-macrophage-markers
Dear Katharina,
the design of the panel really depends on your hypothesis the goal of your experiments. What are you interested to? I mean, if you think that your treatment can alter the APC proporties of monocytes, I definetely recommend CD80, CD86 and MHC-II. If you are interested to check if there is an increase/decrease of immune checkpoints in PBMCs in total, without a focus on a specific subset, I would add CD8 (for cytotoxic T cell), CD56 (for NK cells) and the immune checkpoints Tim-3, KIRs and NKG2A