Ibrutinib pci 32765

The bilateral subcutaneous model was established as for the in vivo anti-cancer efficacy. When the primary tumors reached 100–150 mm³ in volume, mice were injected intratumorally with nMOFs at a dose of 0.11 mg/mouse or PBS. Anti-CD4 (GK1.5, BioXCell, USA), anti-CD8 (OKT-8, BioXCell, USA), mouse IgG (C1.18.4, BioXCell, USA) antibodies Or B cell inhibitor ibrutinib (PCI-32765, Selleckchem) were intraperitoneally injected into the mice (200 μg/mouse) on Day 0 and 5 after the first treatment. Twelve hours post-injection, mice were anesthetized with 2% (v/v) isoflurane, and tumors were irradiated with X-ray at 225 kVp and 13 mA with a 0.3-mm Cu filter. To evaluate the therapeutic efficacy, the tumor growth and body weight were monitored daily.

MCL-RL cells were derived from an MCL patient at the University of Pennsylvania. JeKo-1 and Rec-1 cells were purchased from the American Type Culture Collection (ATCC). SP49 cells were obtained from Dr. Raymond Lai, University of Alberta. Granta-519 cells were purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany). The cell line identity was confirmed by diverse methods including flow cytometry, FISH, and short-tandem repeat (STR) studies. Flow cytometry six-color panel consisted of antibodies against CD45, CD20, CD5, CD23, and immunoglobulin kappa and lambda light chain. FISH detected fusion of immunoglobulin heavy chain (IgH) and cyclin D1 genes. STR test was done at University of Pennsylvania core facility. The cell lines were also found free of mycoplasma contamination using Mycoplasma detection kits from Thermo Fisher (Waltham, MA). All cells were grown in RPMI medium supplemented with 10% fetal bovine serum and 100 μg/ml penicillin/streptomycin under a humidified 37°C/5% CO₂ incubator condition. Ibrutinib (PCI-32765) and CB-839 were purchased from Selleckchem (Houston, TX). [1,6-¹³C₂] glucose, [1,2-¹³C₂] glucose and [U-¹³C₅, U-¹⁵N₂] glutamine were acquired from Cambridge Isotope and Sigma Aldrich.

C57BL/6 mice were obtained from Harlan (Indianapolis, IN) and Jackson Laboratory (Harbor, ME). Il10^–/– mice were obtained from Jackson Laboratory. LPS and anti–β-actin antibody were obtained from Sigma-Aldrich (St. Louis, MO). The polyclonal goat anti-IgM F(ab′)₂ was obtained from ICN/MP Biomedicals (Irvine, CA) or Jackson ImmunoResearch Labs (West Grove, PA). The anti–IL-10R (Clone 1B1.3a) was obtained from BD Biosciences (San Diego, CA). Anti-CD40 (1C10 clone) was a gift from Dr. Maureen Howard and was used as ascites fluid. Syk Inhibitor IV (BAY 61-3606) and SB-203580 were obtained from Calbiochem (San Diego, CA). Ibrutinib (PCI-32765) was obtained from Selleckchem (Houston, TX). Dasatinib was obtained from Bristol-Myers Squibb Company (New York City, NY). Anti–p-Syk, anti-Syk, anti–p-p38 MAPK, anti-p38/MAPK, anti–p-p44/42 MAPK, anti–p-Akt, and anti-Akt antibodies were obtained from Cell Signaling (Danvers, MA). Anti-ERK1 was obtained from Santa Cruz Biotechnology (Santa Cruz, CA).