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Blood, Vol. 111, Issue 2, 517-524, January 15, 2008
The opposite effects of IL-15 and IL-21 on CLL B cells correlate with differential activation of the JAK/STAT and ERK1/2 pathways
Blood de Totero et al.
111: 517
Supplemental Materials for de Totero et al
Files in this Data Supplement:
- Figure S1. IL-15 and IL-21 show opposite effects on CLL cell proliferation (PDF, 56 KB) -
(A)Resting CLL cells were cultured in triplicate wells of 96-well plates in the presence of different concentrations of IL-15 or of IL-21 for 48h and then pulsed with 3H-thymidine (1 µCi/well) for additional 18 hours of culture. DNA synthesis by cells cultured in the presence of IL-21 (50 ng/ml) in combination with IL-15 (50 ng/ml) is also shown.
(B)CD40-activated CLL cells of the same cases above were stimulated with different cytokine concentrations, as in A. The DNA synthesis induced by IL-15 was higher in CD40-activated than in resting cells. IL-21 failed to induce DNA synthesis.
- FigureS2. Comparison of STAT1, 3 and 5, Shc and ERK1/2 signaling pathways -
CLL cells from three cases were stimulated with IL-15 or IL-21 (80 ng/ml) or medium only (CTR) for 45 min, lysed in appropriate buffers and analysed by Western blotting using mAbs specific for phosphorylated forms of STAT1, STAT3, STAT5, Shc and ERK1/2, or for  -actin.
- Figure 3. The Jak inhibitor I blocks IL-21-mediated phosphorylation of STAT1 in CD40-activated CLL cells (PDF, 83 KB) -
Cells were pretreated with Jak inhibitor I (0.4 µM) or with DMSO for 30 min before the addition of IL-21 (80 ng/ml). After additional 10 min of culture cells were processed for Western blot analysis using mAbs specific for tyrosine-phosphorylated STAT1 or for -actin.
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