I noticed in a few papers that after they culture the T-cells with TCR activators and differentiating cytokines for 4-6 days, they wash away the cytokines and stimulate for another day with just the TCR activators. Is this necessary for the cells to produce detectable levels of cytokines? Or would I be able to get good cytokine production without the secondary activation?
Attapong Sinkitjasub
Yes, but it depends on the type of T cells you are working with (e.g., naive vs. effector vs. memory).
Also, the cytokine you are measuring (some cytokines, like IL-2, are transiently expressed early, while others like IFN-γ may require stronger stimulation). Moreover, the initial activation strength (strong initial activation may be sufficient without a secondary boost).
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