Quantitation of rabbit cytokine mRNA by real-time RT-PCR
Quantitation of rabbit cytokine mRNA by real-time RT-PCR
复制标题
DOI：
10.1016/j.cyto.2007.04.002
复制
发表时间：
2007-04-01
期刊：
影响因子：
3.8
通讯作者：
Lukehart, Sheila A.
中科院分区：
文献类型：
--
作者：
Godornes, Charmie;Leader, Brandon Troy;Lukehart, Sheila A.
兔免疫学的基本理解和兔作为疾病模型的使用长期以来一直受到缺乏特异性的免疫学检测这一物种。在本研究中，我们试图开发一种方法来定量兔细胞和组织中的细胞因子表达。我们报告了一种定量实时RT-PCR方法的发展，用于测量兔IFN-γ IL-2，IL-4，IL-10和TNF-α mRNA的相对水平。通过与使用含有相关细胞因子的部分序列的质粒DNA产生的标准曲线进行比较来完成定量。实验研究证明了该测定法定量有丝分裂原刺激后兔脾细胞中细胞因子mRNA水平的适用性。我们还进一步利用该测定来检测实验性梅毒感染期间兔组织中的细胞因子表达。(c)2007爱思唯尔有限公司保留所有权利。
Fundamental understanding of rabbit immunology and the use of the rabbit as a disease model have long been hindered by the lack of immunological assays specific to this species. In the present study, we sought to develop a method to quantitate cytokine expression in rabbit cells and tissues. We report the development of a quantitative real-time RT-PCR method for measuring the relative levels of rabbit IFN-gamma IL-2, IL-4 IL-10 and TNF-alpha mRNA. Quantitation was accomplished by comparison to a standard curve generated using plasmid DNA containing partial sequences of the relevant cytokines. Experimental studies demonstrate applicability of this assay to quantitate cytokine mRNA levels from rabbit spleen cells following mitogen stimulation. We have further utilized this assay to also examine cytokine expression in rabbit tissues during experimental syphilis infection. (c) 2007 Elsevier Ltd. All rights reserved.