Jinliang  Sheng, Chuangfu  Chen, Xia  Yang, Yuanzhi  Wang, Pengyan  Wang, Hui  Zhang, 
    Lipopolysaccharide Stimulus Induces Gene Expression of Cytokines andToll-Like Receptor 2/4 in Ovine Primary Alveolar Macrophages,
    Journal of Animal and Veterinary Advances,
    Volume 10,Issue 24,
    2011,
    Pages 3254-3262,
    ISSN 1680-5593,
    javaa.2011.3254.3262,
    (https://makhillpublications.co/view-article.php?doi=javaa.2011.3254.3262)
    Abstract: 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)-1&#946;, IL-8, Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Interferon (IFN) (&#947;) and Toll-Like Receptor 2/4 (<I>TLR2/4</I>) 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 (C<SUB>T</SUB>) 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-&#947; 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 <I>cytokine</I> gene expression profiles.
    Keywords: Ovine;alveolar macrophage;cytokine;toll-like receptor 2/4;real-time reverse-transcription polymerase chain reaction;China