Spontaneous circulation of myeloid-lymphoid–initiating cells and SCID-repopulating cells in sickle cell crisis
J. Clin. Invest. Christopher E.D. Lamming, et al. 111:811 doi:10.1172/JCI15956 [Go to this article.]

Figure 3
Significantly more LTC-ICs (a), NK-ICs (b), and ML-ICs (c) are present in CD34⁺CD38^–Lin^– cells from AC-SCD patients than in those from NL donors or SS-SCD patients. Single CD34⁺CD38^–Lin^– cells, selected by FACS from normal donors (44–66 wells plated), SS-SCD patients (66–88 wells plated), and AC-SCD patients (66–132 wells plated), were plated in ML-IC assays as previously described (24). After 2 weeks, single CD34⁺CD38^–Lin^– cell progeny were replated in four individual wells, two of which were maintained under LTC-IC and two under NK-IC conditions. For LTC-IC cultures, wells were overlaid after 5 weeks with clonogenic medium, and the presence of CFCs was determined 2 weeks later. NK-IC cells were harvested after 5 weeks and evaluated by FACS for the presence of CD56⁺ NK cells or CD19⁺ B cells. An LTC-IC was determined as a well in LTC-IC cultures where CFCs were present without NK and/or B cells being present in the companion NK-IC cultures. An NK-IC was determined as a well in NK-IC cultures where NK and/or B cells were present without CFCs being present in the companion LTC-IC cultures. An ML-IC was identified when progeny of the initial CD34⁺CD38^–Lin^– cell gave rise to at least one LTC-IC and at least one NK-IC. Significantly more LTC-ICs, NK-ICs, and ML-ICs were present among CD34⁺CD38^–Lin^– cells in AC-SCD blood compared with blood from SS-SCD patients and NL donors. Values shown as 0 represent frequencies below the detection level of the assays. As we plated between 44 and 132 wells, this indicates that fewer than one ML-IC was present among 44–132 CD34⁺CD38^–Lin^– cells. Differences between the groups were evaluated by t test.