I am setting up to test antigen specific T cell responses in patient blood. Some protocols just culture whole PBMCs with antigen (presumably relying on blood monocytes to differentiate into dendritic cells). Others first isolate monocytes and push them to differentiate and the add lymphocytes. Which approach is better?
I also read in few papers they co-culture DC with T lymphocytes that have a separate reason to see specific signalling.
If you just want to see effect of your antigen response by T cells, you may go ahead with whole PBMC culture. Due to stimulation, monocytes will differentiate into DCs and response will be seen effectively.
For to see specific signalling pathway, then you definitely need to co-culture pure populations.
Hi Michael, I was wondering if you used PBMC to activate your antigen-specific T cells? I am planning to carry a similar experiment using synthetic antigens and would love to hear about your experience. Thanks!
Hi Jitendra - Yes, I agree. Thanks
To Ilayda. No, we are looking at PBMC T cells in tumor bearing mice (and patients) to study if they are tumor specific