Cytokine expression in respiratory syncytial virus-infected mice as measured by quantitative reverse-transcriptase PCR

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• 作者：Deng ; Xinqing ; Li ; Haijing ; Tang ; Yi-Wei
• 关键词：RSV ; Real-time PCR ; Cytokines ; Quantitation
• 刊名：Journal of Virological Methods
• 出版年：2003
• 出版时间：February, 2003
• 年：2003
• 卷：107
• 期：2
• 页码：141-146
• 全文大小：522 K

文摘

In the murine model for respiratory syncytial virus (RSV) infection, cytokine patterns induced by vaccinations with either killed (i.e. formalin-inactivated, alum-precipitated) virus (KV) or live virus (LV) have been shown to influence disease expression. To determine the mRNA expression of the cytokines IL-4 and IFN-γ in BALB/c mice challenged with RSV, a real-time quantitative reverse-transcriptase PCR assay was developed. This assay uses 5′-exonuclease fluorogenic probes and is performed on the ABI PRISM 7700 Sequence Detector System (TaqMan). The relative quantitative levels of mRNA for IL-4 and IFN-γ were compared with those measured by an RNase protection assay (RPA) and an enzyme immunoassay (EIA), which are methods used to measure the levels of mRNA and protein, respectively. Results obtained by the TaqMan assay showed that mice primed with KV induces increased IL-4 mRNA production while LV induces increased IFN-γ mRNA, which is in agreement with conventional methods. IL-4 and IFN-γ relative quantities obtained from TaqMan were highly correlated to those determined by RPA (r