Inhibition of NF-kB 1 (NF-kBp50) by RNA interference in chicken macrophage HD11 cell line challenged with Salmonella enteritidis

Abstract

The NF-kB pathway plays an important role in regulating the immunity response in animals. In this study, small interfering RNAs (siRNA) were used to specifically inhibit NF-kB 1 expression and to elucidate the role of NF-kB in the signal transduction pathway of the Salmonella challenge in the chicken HD11 cell line. The cells were transfected with either NF-kB 1 siRNA, glyceraldehyde 3-phosphate dehydrogenase siRNA (positive control) or the negative control siRNA for 24 h, followed by Salmonella enteritidis (SE) challenge or non-challenge for 1 h and 4 h. Eight candidate genes related to the signal pathway of SE challenge were selected to examine the effect of NF-kB 1 inhibition on their expressions by mRNA quantification. The results showed that, with a 36% inhibition of NF-kB 1 expression, gene expression of both Toll-like receptor (TLR) 4 and interleukin (IL)-6 was consistently and significantly increased at both 1 h and 4 h following SE challenge, whereas the gene expression of MyD88 and IL-1β was increased at 1 h and 4 h, respectively. These findings suggest a likely inhibitory regulation by NF-kB 1, and could lay the foundation for studying the gene network of the innate immune response of SE infection in chickens.

Synbiotics for Broiler Chickens—In Vitro Design and Evaluation of the Influence on Host and Selected Microbiota Populations following In Ovo Delivery

“…Gene expression analysis was performed for the selected panel of cytokines: Th1 ( IFNγ , IL12 ), Th2 ( IL4 ), proinflammatory ( IL6 , IL18 , and IL1β ), antiviral ( IFNβ ), and chemokine ( IL8 ). Sequences of primers were based on the literature [19,73–76] or designed based on cDNA nucleotide sequence using NCBI Primer Blast [36] (Tab C in S1 File). …”

Redox Homeostasis in Poultry: Regulatory Roles of NF-κB

“…In order to study the role of NF-κB in the signal transduction pathway of the Salmonella enteritidis -challenged cells, chicken macrophage HD11 cell line and small interfering RNAs (siRNA), specifically inhibiting NF-κB1 expression, were used. In particular, it was found that a 36% inhibition of NF-κB1 expression was associated with increased gene expression of both TLR4 and IL-6 at both 1 h and 4 h following Salmonella challenge [ 274 ]. TLR4 was shown to activate NF-κB signaling during cerebral ischemia–reperfusion, leading to increased secretion of inflammatory cytokines and damage of brain tissue [ 275 ].…”

Effects of hatching system on chick quality, welfare and health of young breeder flock offspring

“…IFNγ can be produced by T helper 1 (Th1) cells and increases the cell-mediated response during an infection, but can be counteracted by IL-10 that has an anti-inflammatory and immune suppressive effect ( Cornelissen et al, 2009 ). NF-κB has a central role in the innate and adaptive immune response toward an infection ( Chiang et al, 2009 ; Oeckinghaus and Ghosh, 2009 ) and activates, amongst others, the production of the cytokine IL-8 ( Liu et al, 2017 ). The lack of difference in transcriptional expression of the selected genes between the 3 hatching systems indicates that with this infection model, the early immune response seems not to be influenced by hatching systems.…”

“…Primers used for the gene expression analysis were extracted from literature and annotation is shown in Table 1 . The 6 genes of interest were claudin, occludin ( Gilani et al, 2018 ), nuclear factors Kappa B (NF-κB) ( Chiang et al, 2009 ). IL8, interferon gamma (IFNγ) ( Cornelissen et al, 2009 ), and IL10 (Genbank accession nr.…”