SUPPLEMENTARY TABLE
Characteristics of the CLL panel and ex vivo response to FLU alone or in combination to Ribavirin
Footnotes
The IC50 were obtained using the MTT assay
Ribavirin sensitization (R value): (FLU IC50) / (IC50 of FLU+ 10μM Ribavirin); R<1 indicates antagonism, R=1 indicates no effect and R>1 indicates sensitization.
Clinical Status; U: Clinically untreated, T: Clinically treated and R: Clinically resistant. Clinical resistance is defined as a relapse within six month of the last treatment
Rai Stage; Rai stage IV represents a more advanced disease than Rai stage 0
IgVH Status; U: unmutated IgVH status and M: mutated IgVH status. The cut off to define IgVH status is 2% mutations respect to germline mutations
CD38, del11 and del17: samples were considered positive if the percentage of positive lymphocytes was ≥ 25%
*Sample isolated from the peripheral blood of a del17 before (#9) and after (#9b) clinical treatment and progression of in the percentage of del17 positive lymphocytes.
SUPPLEMENTARY FIGURE
A) Expression of pAKT (S473) and bcl2 were assessed by western blot in 50 mg of proteins after treatment as indicated for 24 hours in MEC-2 cells.
B) Primary CLL lymphocytes were cultured in the absence (upper panel) or presence of stromal cells (lower panel) and treated with vehicle (red line) or with 10µM Ribavirin (blue line) for twenty four hours. AKT phosphorylation (S473) was assessed by flow cytometry. Labeling with an isotype antibody was used as negative control (black line). Note the shift to the right in basal pAKT (S473) in the presence of stromal cells.
C) Primary CLL lymphocytes were seeded in the presence of a stromal feeder layer. The bars represent the mean value (±SE) of the percentage of AnnexinV positive lymphocytes forty eight hours after in vitro treatment with vehicle, 100 nM BKM-120 or 10 µM Ribavirin (n=4) (Kruskal-Wallis One Way Analysis of Variance on Ranks followed by Mann-Whitney Rank Sum Test cells (**p=0.008, *p=0.01)(left panel).
D) Representative western blot showing the effect of 5mM CGP57380 on peIF4E in two representative M-IgVH and U-IgVH samples.