Our immunization schedule is: 3 s.c. shots of antigen on Alum (d0, d14, d28)
I would like to know if anybody has tested the re-appearance of memory T cells in blood in a time course experiment.
Is there an optimal time point after last immunization for testing T cell reactivity with e.g. an ELISpot assay?
Are the kinetics the same for mice and humans?
we regularily do IFNg ELISPOTs to measure immunogenicityin mice of peptide-based vaccines. We've done time course to determine that 8 days after vaccination is the optimal time to see peak immune responses in the spleen. If your antigens are weak, you could also extract the vaccine-draining lymph node and test them. I've never used mouse PBMC or looked for memory response, but my thinking is that to detect memory you'd need some kind of ex vivo re-stimulation protocol. Also, alum vaccines are biased towards a humoral response, so I don't know what kind of cellular repsonse you could get with that type of vaccine.
Thanks Gen! Is this 8 days optimum achieved after several immunization or only after 1 shot?
The 8 day period is the standard optimum time after a primary immunization (ie after one immunization), when multiple immunizations are applied the maximum response may be observed at a shorter time, as 5 days post the last immuniz dose
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