Any information or any publication about the profiles with these different stimulation methods? such as cell surface markers, cytokines, proliferation rates, cell types stimulated ... I am more interested in human T cell responses.
Also there is stimulation from CEFT peptides. Wondering how to choose the right stimulation method?
There is no right stimulation method. It all depends on your objective and how physiologically relevant you want your stimulation to be. PHA and PMA are polyclonal stimulants ad activate all T cells as long as they are viable. The signaling does not go through the usual MHC-TCR pathway and so if your cells have a defect in these pathways, you wouldn’t see it. So if all you want is a strong positive control for your assay, these would be the way to go. They provide strong stimulation and so you tend to see strong/fast activation, lots of cytokines, fast proliferation and significant activation-induced cell death with time. PHA can also stimulate non-T cells.
aCD3/CD28 (soluble or beads) stimulation is more physiological and is somewhat more representative of TCR signaling. The stimulation is T cell specific but again will activate all viable T cells. If you have a defect in MHC-TCR, you won’t see it but if you have a defect in your intracellular signalling pathways, you would be able to pick it up when using aCD3/CD28. They provide a somewhat strong activation, good amount of cytokines but it takes longer to see proliferation and you have significantly less activation-induced cell death. They are definitely not as powerful as PHA/PMA.
CEF-CEFT pools are the most physiologically relevant activators. The stimulation is T cell specific and it only activates those memory T cells that can respond to CEF/T antigens. So you have significantly less activation, less cytokines and limited proliferation as not all T cells are activated (as compared to when using PHA/PMA/antibodies). But the advantage is that if your cells have defects in TCR-MHC, you will be able to pick it up with antigen specific pools. And with these antigens, you are looking at antigen specific T cell signaling in its entirety.
Hi Julie, thank you for your comprehensive answer. It is very helpful!
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