用TLR4激动剂脂多糖和单磷酰脂质A刺激绵羊和人类血液后的基因表达比较。

Comparison of Gene Expression by Sheep and Human Blood Stimulated with the TLR4 Agonists Lipopolysaccharide and Monophosphoryl Lipid A.

作者信息

Enkhbaatar Perenlei, Nelson Christina, Salsbury John R, Carmical Joseph R, Torres Karen E O, Herndon David, Prough Donald S, Luan Liming, Sherwood Edward R

机构信息

Department of Anesthesiology, the University of Texas Medical Branch, Galveston, TX, United States of America.

Molecular Virology and Microbiology Core, Alkek Center for Metagenomics & Microbiome Research, Baylor College of Medicine, Houston, TX, United States of America.

出版信息

PLoS One. 2015 Dec 7;10(12):e0144345. doi: 10.1371/journal.pone.0144345. eCollection 2015.

BACKGROUND

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).

METHODS

Venous blood, withdrawn from six healthy human adult volunteers (~ 28 years old) and six healthy adult female sheep (~3 years old), was mixed with 30 μL of PBS, LPS (1μg/mL) or MPLA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.

RESULTS

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.

CONCLUSION

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.

背景

能够模拟人类生物学特征的动物模型对于将基础科学发现成功转化为临床实践至关重要。近期对啮齿动物的研究已证明Toll样受体4（TLR4）激动剂作为免疫调节剂在感染模型中的有效性。然而，啮齿动物模型因无法模拟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。

方法

从6名健康成年人类志愿者（约28岁）和6只健康成年雌性绵羊（约3岁）采集静脉血，将其与30μL磷酸盐缓冲液（PBS）、LPS（1μg/mL）或MPLA（10μg/mL）混合，并在室温下于滚动摇床上孵育90分钟。孵育后，将2.5mL血液转移至Paxgene血液RNA管中。使用配备RNA6000纳米实验室芯片的安捷伦生物分析仪进行基因表达分析。用G2565微阵列扫描仪扫描安捷伦基因表达微阵列。鉴定出差异表达的基因。

结果

检测到11431个人类探针和4992个绵羊探针信号高于背景值。其中，1029个人类基因和175个绵羊基因在严格度为1.5倍变化（p<0.05）时差异表达。在这175个绵羊基因中，54个有已知的人类同源基因。在这些基因中，22个在人类样本中的变化超过1.5倍。主要炎症介质基因，如白细胞介素-1（IL-1）、白细胞介素-6（IL-6）和白细胞介素-8（IL-8）、肿瘤坏死因子α（TNFα）、核因子κB（NF-κB）、ETS2、前列腺素内过氧化物合酶2（PTGS2）、五聚体蛋白3（PTX3）、CXC趋化因子配体16（CXCL16）、犬尿氨酸酶（KYNU）和C型凝集素结构域家族4成员E（CLEC4E）在两个物种中均被LPS和MPLA类似地（>2倍）上调。