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Haemophilus parasuis infection activates NOD1/2-RIP2 signaling pathway in PK-15 cells
Bin Ma1, Kexin Hua2, Shanshan Zhou2
1State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
Abstract:
Haemophilus parasuis, an important swine pathogen, was recently proven able to invade into endothelial or epithelial cell in vitro. NOD1/2 are specialized NLRs that participate in the recognition of pathogens able to invade intracellularly and therefore, we assessed that the contribution of NOD1/2 to inflammation responses during H. parasuis infection. We observed that H. parasuis infection enhanced NOD2 expression and RIP2 phosphorylation in porcine kidney 15 cells. Our results also showed that knock down of NOD1/2 or RIP2 expression respectively significantly decreased H. parasuis-induced NF-κB activity, while the phosphorylation level of p38, JNK or ERK was not changed. Moreover, real-time PCR result showed that NOD1, NOD2 or RIP2 was involved in the expression of CCL4, CCL5 and IL-8. Inhibition of NOD1 and NOD2 significantly reduced CCL5 promoter activity, even in a more effective way compared with inhibition of TLR.
Insights
Haemophilus parasuis infection activates NOD1/2 and RIP2 pathways, crucial for swine immune responses. These pathways regulate inflammatory gene expression, offering potential therapeutic targets for H. parasuis infections.
Area of Science:
- Immunology
- Microbiology
- Cell Biology
Background:
- Haemophilus parasuis is a significant swine pathogen capable of intracellular invasion.
- NOD1 and NOD2 (nucleotide-binding oligomerization domain-containing proteins 1 and 2) are key intracellular pattern recognition receptors involved in detecting invasive pathogens.
Purpose of the Study:
- To investigate the role of NOD1/2 in mediating inflammatory responses during Haemophilus parasuis infection in porcine cells.
Main Methods:
- Porcine kidney 15 cells were infected with H. parasuis.
- NOD1/2 and RIP2 expression and phosphorylation were analyzed.
- NF-κB activity and MAPK signaling pathways (p38, JNK, ERK) were assessed.
- Quantitative real-time PCR and promoter activity assays were performed.
Main Results:
- H. parasuis infection increased NOD2 expression and RIP2 phosphorylation in porcine kidney cells.
- Knockdown of NOD1/2 or RIP2 significantly reduced H. parasuis-induced NF-κB activity.
- NOD1, NOD2, and RIP2 were involved in the expression of inflammatory chemokines CCL4, CCL5, and IL-8.
- Inhibition of NOD1 and NOD2 more effectively reduced CCL5 promoter activity than TLR inhibition.
Conclusions:
- NOD1/2 and RIP2 play a significant role in the inflammatory response to H. parasuis infection.
- The NOD1/2-RIP2 pathway is critical for NF-κB activation and the expression of specific inflammatory mediators during H. parasuis infection.
- NOD1/2 signaling is a key component of the innate immune response against H. parasuis, potentially more critical than TLRs for CCL5 induction.
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