TY - JOUR T1 - Lipopolysaccharide Stimulus Induces Gene Expression of Cytokines andToll-Like Receptor 2/4 in Ovine Primary Alveolar Macrophages AU - Sheng, Jinliang AU - Chen, Chuangfu AU - Yang, Xia AU - Wang, Yuanzhi AU - Wang, Pengyan AU - Zhang, Hui JO - Journal of Animal and Veterinary Advances VL - 10 IS - 24 SP - 3254 EP - 3262 PY - 2011 DA - 2001/08/19 SN - 1680-5593 DO - javaa.2011.3254.3262 UR - https://makhillpublications.co/view-article.php?doi=javaa.2011.3254.3262 KW - Ovine KW -alveolar macrophage KW -cytokine KW -toll-like receptor 2/4 KW -real-time reverse-transcription polymerase chain reaction KW -China AB - In this study, Real-Time Reverse-Transcription Polymerase Chain Reaction (RT-PCR) assays were performed with EvaGreen to investigate the dynamics of cytokine (Interleukin (IL)-1β, IL-8, Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Interferon (IFN) (γ) and Toll-Like Receptor 2/4 (TLR2/4) gene expression in ovine primary Alveolar Macrophages (AMs) following Lipopolysaccharide (LPS) stimulation. Expression of cytokine and TLR2/4 mRNA was quantified by comparison of Cycle threshold (CT) values with a standard curve generated from plasmid DNA containing the target gene. Examination of LPS-stimulated ovine AMs revealed that cytokine mRNA expression peaked between 4 and 12 h with the exception of IFN-γ mRNA which peaked around 16 h post stimulation. Furthermore, TLR2 and TLR4 mRNA expression rapidly increased post-stimulation and peaked 20 min post-stimulation at a level which was maintained throughout the procedure. In summary, a sensitive and reliable real-time RT-PCR protocol was implemented for the analysis of ovine TLR2/4 and cytokine gene expression profiles. ER -