Dear All,
I am interested in analysing the expression of the following exhaustion markers : PD-1, CTLA-4 , TIM-3 and LAG-3 on Black 6 mouse naive splenocytes using flow cytometry. I am in the process of titrating my antibodies for these specific markers and I need to first activate my cells to upregulate those markers.
But I am not sure which activation / stimulation procedure would be best to ensure the upregulation of these markers. Is CD3/CD28 beads or Con A better? Knowing they act through different mechanisms. And for how long should I keep my cells in culture to ensure maximal activation of my cells?
Thank you very much for your precious input!
Those aren't exactly "exhaustion" markers, so pushing for T cell activation won't be that efficient. The best way, i think is to change the cytokine environment. So... the below is a link to a paper for dexamethasone... There should be others... hopefully someone else will have a better idea.
Article Dexamethasone enhances programmed cell death 1 (PD-1) expres...
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