Autocrine secretion of soluble factors protects CLL cells from TLR9-mediated apoptosis. (A) Viability of Zap-70–positive CLL cells cultured with or without DSP30 for 72 hours. Line represents the mean ± SD of n = 3 prosurvival responder patient samples (left). Viability of ZAP-70–negative CLL cells cultured with or without DSP30 for 72 hours in fresh medium or in the respective conditioned media (CM) derived from Zap-70–positive samples (n = 3 for CM control and CM DSP30). Data are presented as the mean ± SD of n = 4 patient samples in a bar diagram (right panel). (B) Cytokines secreted by purified CLL cells, cultured with or without DSP30 for 72 hours, quantified by enzyme-linked immunosorbent assay (ELISA). Data are presented in a bar diagram as the mean ± SEM of n = 20 (interleukin [IL] 2), n = 12 (IL-15), and n = 28 patient samples for IL-21, BAFF, and IL-10. (C) DSP30-mediated changes in the expression of glycoproteins secreted by purified Zap-70–positive CLL assessed after 72 hours of culture. Results of mass spectrometric analysis are presented as the mean of n = 3 patients samples in a volcano plot. P values were calculated with Student t test for all proteins based on the log2 intensity ratios. Proteins with P ≤ .05 were considered as hits. (D) sIgM produced by CLL cells cultured with or without DSP30 for 48 hours, quantified by ELISA. Data are presented in a bar diagram as the mean ± SEM of n = 24 patient samples. P values for the entire figure are indicated by asterisks: ****P < .0001; ***P < .001; **P < .01; *P < .05.