I am setting up an experiment to characterise tetanus-specific T-cells in humans. I try to decide if it is best to use whole tetanus toxoid or peptide pools.

I want to use CD154 to detect Ag-specific CD4 Tcell following stimulation of PBMC with tetanus toxoid. I understand that CD154 is relatively early marker (6 hours) but I cannot reconcile the use of whole protein and such an early time point .... how a whole protein can be processed and presented in such a short time ? The objective being to assess TCRseq and transcriptomic data on tetanus specific T-cell, would you suggest changing my activation marker or use peptide pools even if we will probably restrict the TCR breath of the population obtained?

Many hours of reading to realise that I could not find any consensus on the question with time of stimulation varying fro a few hours to several days with whole protein or peptide pools .... I look forward for your opinions!