hey...I m planing to activate human T cells after asolated from PBMCs and before, so I know some about PHA that can activate T cells. also the compaine between PMA and Ionomycin also can activate them. so if there any help in this you can brovide me that will be so Kind and helpful.
I plane to measure the Granzymes that secreted by T cells.
Hi, the "classical" way to activated CD3+ CD4+ CD25- cells are indeed by treating cells with a cocktail of PHA and IL2 (2mM both) or by treating cells with anti CD3 and CD28 (commercial Kit). You have to wait at least 3h after the purification to begin the treatment.
I'd say the best way to activate T-cells would be using anti-CD3 and anti-CD28 antibodies, followed by supplementing the medium with IL2. Alternatively, you can also activate them using superantigens like concanavalin A (conA). The advantage of using concanavalin A is that you don't need to coat the wells unlike anti-CD3 and anti-CD28 antibodies. Instead you can directly add conA to your cells and you're good to go from there. But make sure you wash the cells off conA before proceeding with any kind of treatment on your cells.
Hello! Your doubt is about specific stimulus, right? I Think that you would use PMA . I'm going to send for you one recent article about this it.
http://www.jimmunol.org/content/early/2011/11/14/jimmunol.1100891.full.pdf
Good luck!!!
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