Original Article
Copyright ©2013 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Biol Chem. May 26, 2013; 4(2): 18-29
Published online May 26, 2013. doi: 10.4331/wjbc.v4.i2.18
Refractoriness of interferon-beta signaling through NOD1 pathway in mouse respiratory epithelial cells using the anticancer xanthone compound
Zaifang Yu, Jarrod D Predina, Guanjun Cheng
Zaifang Yu, Jarrod D Predina, Guanjun Cheng, Thoracic Oncology Research Laboratory, 1016B ARC, University of Pennsylvania, Philadelphia, PA 19104, United States
Zaifang Yu, Jarrod D Predina, Guanjun Cheng, Department of Surgery, Thoracic Surgery Research Laboratory, University of Pennsylvania, Philadelphia, PA 19104, United States
Author contributions: Cheng G drafted manuscript; Yu Z and Predina JD revised manuscript.
Supported by NCI PO1 CA 66726
Correspondence to: Dr. Guanjun Cheng, Thoracic Oncology Research Laboratory, 1016B ARC, University of Pennsylvania, 3615 Civic Center Boulevard, Philadelphia, PA 19104, United States. guanjun@mail.med.upenn.edu
Telephone: +1-215-5732593 Fax: +1-215-5734469
Received: January 14, 2013
Revised: March 19, 2013
Accepted: April 28, 2013
Published online: May 26, 2013
Processing time: 116 Days and 9.1 Hours
Abstract

AIM: To explore the possibility that nucleotide oligomerization domain 1 (NOD1) pathway involved in refractoriness of interferon-β signaling in mouse respiratory epithelial cells induced by the anticancer xanthone compound, 5,6-dimethylxanthenone-4-acetic acid (DMXAA).

METHODS: C10 mouse bronchial epithelial cells were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine, 100 units/mL penicillin, 100 g/mL streptomycin. Pathogen-free female BALB/c mice were used to explore the mechanisms of refractoriness of interferon-signaling. Mouse thioglycollate-elicited peritoneal macrophages, bone marrow derived macrophages and bone marrow derived dendritic cells were collected and cultured. The amount of interferon (IFN)-inducible protein-10 (IP10/CXCL10), macrophage chemotactic protein (MCP1/CCL2) and interleukin (IL)-6 secreted by cells activated by DMXAA was quantified using enzyme-linked immunosorbent assay kits according to the instructions of the manufacturers. Total RNA was isolated from cells or nasal epithelium with RNeasy Plus Mini Kit, and cDNA was synthesized. Gene expression was checked using Applied Biosystems StepOne Real-Time Polymerase Chain Reaction System. Transfection of small interfering RNA (siRNA) control, NOD1 duplexed RNA oligonucleotides, and high-mobility group box 1/2/3 (HMGB1/2/3) siRNA was performed using siRNA transfection reagent.

RESULTS: DMXAA activates IFN-β pathway with high level of IFN-β dependent antiviral genes including 2’, 5’-oligoadenylate synthetase 1 and myxovirus resistance 1 in mouse thioglycollate-elicited peritoneal macrophages, bone marrow derived macrophages and bone marrow derived dendritic cells. Activation of IFN-β by DMXAA involved in NOD1, but not HMGB1/2/3 signal pathway demonstrated by siRNA. NOD1 pathway plays an important role in refractoriness of IFN-β signaling induced by DMXAA in mouse C10 respiratory epithelial cells and BALB/c mice nasal epithelia. These data indicate that DMXAA is not well adapted to the intrinsic properties of IFN-β signaling. Approaches to restore sensitivity of IFN-β signaling by find other xanthone compounds may function similarly, could enhance the efficacy of protection from influenza pneumonia and potentially in other respiratory viral infections.

CONCLUSION: NOD1 pathway may play an important role in refractoriness of IFN-β signaling in mouse respiratory epithelial cells induced by DMXAA.

Keywords: Innate immunity; Interferon; Refractoriness; Xanthone; Bronchial epithelium

Core tip: We recently demonstrated that a small, cell-permeable compound, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), was able to induce production of interferon (IFN)-γ and IFN-β-dependent proteins and protect epithelial cells in vitro from virally-induced cell death and to protect mice from a lethal dose of H1N1 influenza A virus. DMXAA that activates multiple antiviral pathways including IFN-β pathway is an attractive strategy in antiviral therapies. Nucleotide oligomerization domain 1 pathway may play an important role in refractoriness of IFN-β signaling in mouse respiratory epithelial cells induced by DMXAA.