Comparison of Gene Expression by Sheep and Human Blood Stimulated with the TLR4 Agonists Lipopolysaccharide and Monophosphoryl Lipid A.
Comparison of Gene Expression by Sheep and Human Blood Stimulated with the TLR4 Agonists Lipopolysaccharide and Monophosphoryl Lipid A.
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DOI：
10.1371/journal.pone.0144345
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Sherwood ER
中科院分区：
文献类型：
--
作者：
Enkhbaatar P;Nelson C;Salsbury JR;Carmical JR;Torres KE;Herndon D;Prough DS;Luan L;Sherwood ER
模仿人类生物学的动物模型对于成功地将基础科学发现转化为临床实践非常重要。最近在啮齿动物中的研究已经证明了TLR4激动剂在感染模型中作为免疫调节剂的功效。然而，啮齿动物模型因不能模仿人类对微生物产物免疫反应的重要特征而受到批评。本研究的目的是比较人类和绵羊血液对TLR4激动剂脂多糖（LPS）和单磷酰脂质A （MPLA）的基因组反应。取6名健康成年志愿者（~ 28岁）和6只健康成年母羊（~3岁）的静脉血，分别与30 μL PBS、LPS （1μg/mL）或MPLA （10μg/mL）混合，室温下摇床孵育90分钟。孵育后取2.5 mL血液转入Paxgene blood RNA管。基因表达分析使用安捷伦生物分析仪和RNA6000纳米实验室芯片进行。用G2565微阵列扫描仪扫描安捷伦基因表达微阵列。鉴定出差异表达基因。在背景上方检测到11,431个人类探针和4,992个绵羊探针。其中人基因1029个，羊基因175个，差异表达幅度为1.5倍（p<0.05）。在这175个绵羊基因中，有54个已知与人类同源。在这些基因中，22个在人类样本中发生了1.5倍的变化。主要炎症介质IL-1、IL-6和IL-8、TNF α、NF-kappaB、ETS2、PTGS2、PTX3、CXCL16、KYNU和cle4e的基因在LPS和MPLA的作用下同样上调（bbbb2倍）。绵羊外周血对LPS和MPLA的基因组反应与在人类中观察到的非常相似，支持将羊模型用于模拟人类炎症疾病的转化研究和基于tlr的免疫调节剂的研究。
Animal models that mimic human biology are important for successful translation of basic science discoveries into the clinical practice. Recent studies in rodents have demonstrated the efficacy of TLR4 agonists as immunomodulators in models of infection. However, rodent models have been criticized for not mimicking important characteristics of the human immune response to microbial products. The goal of this study was to compare genomic responses of human and sheep blood to the TLR4 agonists lipopolysaccharide (LPS) and monophosphoryl lipid A (MPLA). Venous blood, withdrawn from six healthy human adult volunteers (~ 28 years old) and six healthy adult female sheep (~3 years old), was mixed with 30 μL of PBS, LPS (1μg/mL) or MPLA (10μg/mL) and incubated at room temperature for 90 minutes on a rolling rocker. After incubation, 2.5 mL of blood was transferred to Paxgene Blood RNA tubes. Gene expression analysis was performed using an Agilent Bioanalyzer with the RNA6000 Nano Lab Chip. Agilent gene expression microarrays were scanned with a G2565 Microarray Scanner. Differentially expressed genes were identified. 11,431 human and 4,992 sheep probes were detected above background. Among them 1,029 human and 175 sheep genes were differentially expressed at a stringency of 1.5-fold change (p<0.05). Of the 175 sheep genes, 54 had a known human orthologue. Among those genes, 22 had > 1.5-fold changes in human samples. Genes of major inflammatory mediators, such as IL-1, IL-6 and IL-8, TNF alpha, NF-kappaB, ETS2, PTGS2, PTX3, CXCL16, KYNU, and CLEC4E were similarly (>2-fold) upregulated by LPS and MPLA in both species. The genomic responses of peripheral blood to LPS and MPLA in sheep are quite similar to those observed in humans, supporting the use of the ovine model for translational studies that mimic human inflammatory diseases and the study of TLR-based immunomodulators.