Purification of Primary CLL and Normal B-cells

Blood samples from a total of 67 previously untreated CLL patients and 30 age-matched, normal healthy subjects were obtained for this study. Samples from normal individuals (N) or CLL patients (P) were assigned arbitrary numbers and presented in Supplementary Table S1. The mean age for the 67 CLL patients was 67.7 years (standard deviation of 10.0, range 44–91 years). Among the CLL patients 73% (49/67) were male, 22% (14/65) were Rai Stage III or IV, 63% (29/46) had IGHV mutation, 30% (15/50) ZAP70 positive (≥20%), 27% (16/60) CD38 positive ((≥20%), and 23% (14/61) normal FISH. CLL samples were randomly selected for each experiment based on cell availability but not based on any patient characteristics or experiment outcomes. All patients provided written informed consent according to the Declaration of Helsinki to the OUHSC and Mayo Clinic Institutional Review Boards, which approved these studies.
Primary CLL cells or normal B-cells were purified from blood of CLL patients or healthy individuals, respectively, using RosetteSep B-cell enrichment kit (STEMCELL)^{20 (link),21 (link)}. Typical purification range of CD5⁺/CD19⁺ CLL cells was ≥95–99% and >95% pure CD19⁺ B-cells were obtained from normal blood.