What is the best timing to pulse dendritic cell vaccines with TAAs-peptides? How many hours before inducing maturation?
I have also noticed some conduct peptide pulsing during the process of monocyte-to-DC differentiation (e.g. day 3).
Thanks for any answers!
Never before maturation. Pulse DC a few hours before vaccination to prevent MHC/peptide recyling. Proteins were added to immature DC, peptides only after terminal maturation.
Regards,
Helmut
Dear prof. Jonuleit,
thank you for your answer! Do you believe that pulsing the DCs with whole TAA proteins could be more beneficial than using peptides alone? Could such an approach potentially lead to loading of both MHC I and MHC II, therefore facilitating CD4+ helper response, which was recently shown to be important in e.g. melanoma?
Sincerely,
U. Svajger
Dear Urban,
It's dependent on your project. Tumoe-associated proteins allows antigen-loading independent on individual MHC haplotypes of individual patients.
Peptides are optimal if specific MHC/peptide restricted T cells should be investigated.
Regards,
Helmut
As mentioned above. Need to pulse immature DC. Just be careful for pulsing immature DC as it is difficult to define it. Transfer from one well to another would already compromise the immature status. Mature factors are also important. Pulsing different type of protein is another you should consider. As far as I know, long peptide enclose defined short epitope is better than whole protein in term of potency both for class I or class II.
After 4-days, pulse immature DC, then allow at least 6 hours before maturation
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