Activation of CD160 triggers IL-6 and IL-8 secretion in CLL cells. (A) Cells (5 × 105) in 100 μL of medium were incubated with the isotype control and either CL1-R2 or Pan-Ig antibody (anti-IgG,M,A). The human CBA Flex kit was used for simultaneous measurement of IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α, and VEGF expression by a FACSCanto after 24-hour incubation. (Top panel) Complete results for 1 case (CLL15) are shown. Pan-Ig and CL1-R2 significant production of IL-6 (*P = .023, **P = .019) and IL-8 (*P = .015, **P = .045) versus control conditions. (Bottom panel) The median and range of cytokine release for all cases are shown (IL-8, n = 5; other cytokines, n = 10). (B) Fold increase of IL-6 and IL-8 compared with control conditions for all cases (IL-8, n = 5; IL-6, n = 10). (C) Cells (5 × 105) in 100 μL of medium were pretreated with 10μM or 20μM LY249002 for 1 hour and then incubated with isotype control or CL1-R2 for 24 hours. IL-6 expression was detected with the CBA Flex kit. The data shown are the mean ± SEM from 3 patients; *P = .014, **P < .001 comparing CL1-R2 alone versus LY294002 followed by CL1-R2.
