Phenotypical analysis by flow cytometry
After the incubation period, specific proliferations were assessed by
flow cytometry in a FACS Canto II cytometer (BD Biosciences Milpitas,
USA) analysing the CFSEdim expression in
CD3+, CD4+, CD8+and NK cells and in different subpopulations:
CD3+CD4+CXCR3+IFNγ+(CD4+Th1);
CD3+CD4+CRTH2+IL-4+(CD4+Th2)35; NK cells including the
subtypes,
CD3-CD56+Perforin+(NKPerf) and
CD3-CD56+IFNγ+(NKIFN-γ).11 Data were analysed by
FlowJo software (BD Biosciences Milpitas, USA) following different gate
strategies (Figure S1B). Results were expressed as Proliferation Index
(PI) calculated as (%CFSEdim stimulated
lymphocytes+moDCs-%CFSEdim unstimulated
lymphocytes+moDCs)/ %CFSEdimlymphocytes.35