Interleukin 4 (il 4)

Ibrutinib was purchased from Selleckchem (Houston, TX, USA), cerdulatinib was provided by Portola Pharmaceuticals Inc. (South San Francisco, CA, USA), CpG (ODN2006, stimulatory CpG-ODN type B, human specific) was purchased from Invivogen (San Diego, CA, USA), IL-6 was from R&D Systems (Minneapolis, MN), and IL-4 and CD40L were from Enzo Life Sciences (Plymouth Meeting, PA, USA). Antibodies: anti-phosphorylated BTK (p-BTK) (Y223), p-IκBα (S32/36) p-STAT3 (Y705), STAT3, MCL-1, p-JAK1 (Y1022), p-JAK3 (Y980), p-STAT6 (Y641), p-STAT3 (S727), and p-JAK2 (T1007/1008) were purchased from Cell Signaling Technology (Danvers, MA, USA); anti-total BTK antibody, poly ADP-ribose polymerase (PARP), and the BrdU detection kit were from BD Biosciences (San Jose, CA, USA); anti-p65, STAT-6, STAT-3, JAK1, JAK2, JAK3 and GAPDH antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). For flow cytometry, FITC-anti-CD19 (clone HIB19) and PE-anti-CD5 (clone UCHT2) were purchased from eBioscience (San Diego, CA, USA). Alexa Fluor® 647-anti-p-AKT (Ser473) and Alexa Fluor® 488 anti-p-p44/42 MAPK (ERK1/2) (T202/Y204) were purchased from Cell Signaling Technology, and PE-anti-p-PLCγ2 (Y759) was purchased from BD Bioscience.

The serum cytokine and chemokine levels were measured using a mouse ELISA kit, according to the manufacturer’s instructions (IL-4 and IL-6, Enzo Life Sciences, Farmingdale, NY, USA; TSLP, MCP-1, Elabscience, Huston, ID, USA; IL-1β, TARC, MDC, and RANTES, R&D Systems Inc., Minneapolis, MN, USA).