Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5589190 | Gene | 2017 | 8 Pages |
Abstract
Cluster of differentiation antigen 14 (CD14) is the membrane receptor protein in Toll-like Receptor 4 (TLR4) signaling pathway, which plays an important regulation role in not only innate immune response but also adaptive immune response. In this study, the pig kidney epithelial cell (PK15) line with CD14 gene silencing mediated by lentivirus was established and cells of CD14-RNAi and NC group were exposed to three kinds of Escherichia coli (E. coli F18ab, E. coli F18ac and E. coli K88ac) and LPS. Then qPCR and western blot were used to detect expression levels of TLR4 signaling pathway-related genes. Finally, ELISA was used to detect the level of proinflammatory cytokines in the cell culture supernatant. The results showed that the expression level of TLR4 signaling pathway-related genes in the entire signal pathway had obvious increases when cells were exposed to the stimulation induced by E. coli and LPS. In addition, the expression levels of CD14-RNAi group were overall significantly lower than NC group (PÂ <Â 0.05 or PÂ <Â 0.01), which was the same with the release levels of proinflammatory cytokines. This study revealed that pig CD14 gene silencing partially inhibited immune response to E. coli F18 invasion mediated by TLR4 signaling pathway.
Keywords
IL-1βmembrane CD14myeloid differentiation proteinSYBRmCD14MD2sCD14TLR4CD14LTAPRRGPiATCCE. coliLBPIFN-αMYD88LPSqPCRFBSDMEMDulbecco's modified Eagle Mediumlipoteichoic acidEscherichia coliinterferon-alphaInterleukin-1βtumor necrosis factor-alphaGene silencingPigfetal bovine serumTNF-αlipopolysaccharidemajor histocompatibility complexMHCAmerican Type Culture CollectionSoluble CD14quantitative real-time PCRImmune responseLPS-binding proteinmyeloid differentiation protein 2Toll-like receptor 4PRR, Pattern recognition receptor
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Authors
Chaohui Dai, Haifei Wang, Guoqiang Zhu, Shenglong Wu, Wenbin Bao,