Absolute values T and NK cells according to the iNKT/T ratios, origin, and functionality of iNKT cells. (A) PBMCs from patients were harvested at 3 time points after HSCT and means (± SD) of absolute numbers of iNKT, T (CD4+ or CD8+), and NK (CD3−CD56+) cells are represented. The numbers of analyzed patients per time point were 25 on day 30, 23 on day 60, and 18 on day 90 for those with an iNKT/T ratio ≥ 10−3 and 42 on day 30, 43 on day 60, and 42 on day 90 for those with an iNKT/T ratio < 10−3. Although patients with an iNKT/T ratio ≥ 10−3 had also greater iNKT circulating cells in absolute numbers, they had similar T and NK reconstitution during the first 3 months after HSCT independent of their iNKT/T ratio (P < .0001, Mann-Whitney test). (B) CD4+ T or iNKT cells obtained after in vitro expansion were electronically sorted and genomic DNA extracted. In all patients analyzed, both CD4+ T and iNKT cells had a donor genotype. (C) Both CD4+ and CD4− iNKT cell subsets from patients independently of their iNKT/T ratios were expanded in culture before their electronic sorting. These cells were then stimulated and the levels of IL-4 and IFN-γ measured in the supernatants demonstrate their functionality. NS indicates nonsignificant.
