T cell enrichment
Using Sepax C-pro, the fresh leukopak was washed and concentrated using
CliniMACS PBS/EDTA Buffer (Miltenyi Biotech) supplemented with 0.1% HSA
and then incubated with antibodies specific to CD4 and CD8 (Miltenyi
Biotec) at room temperature for 30 minutes. Labeled leukopak cells were
then transferred on CliniMACS Plus (Miltenyi Biotech) and positive
fraction of flow through was collected according to manufacturers’
instructions.