I have been trying to culture antigen specific specific T cell culture from PBMC's. I stumulate the PBMSC's with peptides at 1ug/ml dose, followed by 5ng/ml IL2. Thevpeptide specific T cell growth is not great compared to Immunocult stimulation. Would addition of either IL-15 or IL-7 improve T cell growth? Which interleukin is better and what would be the ideal concentration to use?
Of course proliferation using peptides will be a lot less than using immunocult, since the latter will stimulate ALL the t cells. And you don't have to deal with presentation of the peptide by APCs.
Just extend the time in culture. Or use differentiated macrophages loaded with peptide.
Shen-An Hwang is right. You can't expect the same level of stimulation as immunocult, when using antigen. How long are you stimulating the cells for? What is your readout? What do you mean by “followed by IL-2”?
I used to add IL-2 along with the peptides. I also did a study comparing IL-7 and IL-15 supplementation (on top of IL-2). The supplementation did improve T cell responses. Since I couldn’t find a difference between the two at least in my experimental setup, I decided to stick with IL-2 and IL-7. I can't remember the exact conc. I used but 10 ng/mL should work.
Thanks@ Shen-An Hwang and @ Julie Joseph. Extending the time in culture seems to be working..
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