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Refractoriness of interferon-beta signaling through NOD1 pathway in mouse respiratory epithelial cells using the
Zaifang Yu1, Jarrod D Predina, Guanjun Cheng
1Zaifang Yu, Jarrod D Predina, Guanjun Cheng, Thoracic Oncology Research Laboratory, 1016B ARC, University of Pennsylvania, Philadelphia, PA 19104, United States.
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.
Insights
The nucleotide oligomerization domain 1 (NOD1) pathway is involved in interferon-β (IFN-β) signaling refractoriness induced by the anticancer compound DMXAA in mouse respiratory cells. This suggests potential strategies to enhance IFN-β signaling for treating respiratory viral infections.
Area of Science:
- Immunology
- Cell Biology
- Pharmacology
Background:
- Interferon-β (IFN-β) signaling is crucial for antiviral responses.
- The anticancer compound 5,6-dimethylxanthenone-4-acetic acid (DMXAA) activates the IFN-β pathway.
- Understanding mechanisms of IFN-β signaling refractoriness is important for therapeutic applications.
Purpose of the Study:
- To investigate the role of the nucleotide oligomerization domain 1 (NOD1) pathway in DMXAA-induced IFN-β signaling refractoriness.
- To explore the potential of modulating the NOD1 pathway for enhancing IFN-β signaling efficacy.
Main Methods:
- Utilized C10 mouse bronchial epithelial cells and BALB/c mice models.
- Quantified cytokine secretion (IP10/CXCL10, MCP1/CCL2, IL-6) using ELISA.
- Assessed gene expression via Real-Time Polymerase Chain Reaction (RT-PCR) after siRNA-mediated knockdown of NOD1 and HMGB1/2/3.
Main Results:
- DMXAA activated IFN-β-dependent antiviral genes in various immune cells.
- siRNA experiments demonstrated that NOD1, but not HMGB1/2/3, mediated DMXAA's activation of IFN-β signaling.
- The NOD1 pathway was identified as a key player in DMXAA-induced IFN-β signaling refractoriness in respiratory epithelial cells and nasal epithelia.
Conclusions:
- The NOD1 pathway significantly contributes to the refractoriness of IFN-β signaling induced by DMXAA in mouse respiratory epithelial cells.
- These findings suggest that targeting the NOD1 pathway could restore IFN-β signaling sensitivity.
- Modulating IFN-β signaling may enhance therapeutic efficacy against influenza pneumonia and other respiratory viral infections.
