Buffy Coat PBMC Isolation
Donor PBMC were isolated from healthy donor buffy coats by ficoll-paque (GE Healthcare) density centrifugation (450g for 40 minutes) to set up flow cytometry panels (surface phenotype and intracellular cytokine assay). Donor PBMC from n=6 buffy coats were also characterised for cytokine expression in relation to T cell memory markers (see intracellular cytokine assay).