PCR primers and probes were designed based on the sequences reported in GenBank with the Primer Express software version 1.2 (Applied Biosystems) as follows: FOXP3 forward primer, 5′-GAAACAGCACATTCCCAGAGTTC-3′; FOXP3 reverse primer, 5′-ATGGCCCAGCGGATGAG-3′; EF-1a forward primer, 5′-CTGAACCATCCAGGCCAAAT-3′; and EF-1a reverse primer, 5′-GCCGTGTGGCAATCCAAT-3′, as previously described (Mantel et al., 2007).