绵羊细粒棘球绦虫虫卵感染早期发生即时固有免疫反应：来自微阵列分析的证据
An Immediate Innate Immune Response Occurred In the Early Stage of E. granulosus Eggs Infection in Sheep: Evidence from Microarray Analysis.
作者信息
Hui Wenqiao, Jiang Song, Tang Jishun, Hou Hongyan, Chen Sheng, Jia Bin, Ban Qian
机构信息
Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agriculture Sciences, Road Nongkenan, Hefei, 230031, Anhui, People's Republic of China; College of Animal Science and Technology, Shihezi University, Road Beisi, Shihezi 832003, Xinjiang, People's Republic of China.
College of Animal Science and Technology, Shihezi University, Road Beisi, Shihezi 832003, Xinjiang, People's Republic of China.
出版信息
PLoS One. 2015 Aug 7;10(8):e0135096. doi: 10.1371/journal.pone.0135096. eCollection 2015.
BACKGROUND
Cystic Echinococcosis(CE), caused by infection with the larval stage of the cestode Echinococcus granulosus (E. granulosus), is a chronic parasitic zoonosis, with highly susceptible infection in sheep. However, the comprehensive molecular mechanisms that underlie the process of E. granulosus infection in the early stage remain largely unknown. The objective of this present study was to gain a cluster of genes expression profiles in the intestine tissue of sheep infected with CE.
METHODS
Nine healthy sheep were divided into infection group and healthy controls, with six infected perorally 5000 E. granulosus eggs suspended in 1000 μl physiological saline and three controls perorally injected 1000 μl physiological saline. All animals were sacrificed at 4 hours post-infection, respectively. The intestine tissue was removed and the RNA was extracted. In the infection group, the biology replicates were designed to make sure the accuracy of the data. The ovine microarrays were used to analyze changes of gene expression in the intestine tissue between CE infected sheep and healthy controls. Real-time PCR was used to assess reliability of the microarray data.
RESULTS
By biology repeats, a total of 195 differentially expressed genes were identified between infected group and controls at 4 hours post-infection, with 105 genes related to immune responses, while 90 genes associated with functions including energy metabolism, fat soluble transport, etc. Among the 105 immunity genes, 72 genes showed up-regulated expression levels while 33 showed down-regulation levels. Function analysis showed that most of up-regulated genes were related to innate immune responses, such as mast cell, NK cell, cytokines, chemokines and complement. In addition, Real-time PCR analysis of a random selection of nine genes confirmed the reliability of the microarray data.
CONCLUSION
To our knowledge, this is the first report describing gene expression profiles in the intestine tissue of CE infection sheep. These results suggested that the innate immune system was activated to elicit immediate defense in the intestine tissue where E. granulosus invaded in at 4 hour-post infection. Furthermore, future interest will also focus on unraveling similar events, especially for the function of adaptive immunity, but at late stage infection.
背景
囊型包虫病（CE）是由细粒棘球绦虫（E. granulosus）幼虫感染引起的一种慢性寄生虫人畜共患病，绵羊对此高度易感。然而，细粒棘球绦虫早期感染过程的综合分子机制仍 largely 未知。本研究的目的是获得感染 CE 的绵羊肠道组织中的一组基因表达谱。
方法
将 9 只健康绵羊分为感染组和健康对照组，6 只经口感染悬浮于 1000 μl 生理盐水中的 5000 个细粒棘球绦虫虫卵，3 只对照组经口注射 1000 μl 生理盐水。所有动物分别在感染后 4 小时处死。取出肠道组织并提取 RNA。在感染组中，设计生物学重复以确保数据的准确性。使用绵羊微阵列分析 CE 感染绵羊和健康对照组肠道组织中基因表达的变化。实时 PCR 用于评估微阵列数据的可靠性。
结果
通过生物学重复，在感染后 4 小时，感染组和对照组之间共鉴定出 195 个差异表达基因，其中 105 个基因与免疫反应相关，90 个基因与能量代谢、脂溶性转运等功能相关。在 105 个免疫基因中，72 个基因表达上调，33 个基因表达下调。功能分析表明，大多数上调基因与先天免疫反应相关，如肥大细胞、NK 细胞、细胞因子、趋化因子和补体。此外，对随机选择的 9 个基因进行实时 PCR 分析证实了微阵列数据的可靠性。
结论
据我们所知，这是第一份描述 CE 感染绵羊肠道组织中基因表达谱的报告。这些结果表明，在感染后 4 小时，细粒棘球绦虫侵入的肠道组织中先天免疫系统被激活以引发即时防御。此外，未来的研究兴趣还将集中在揭示类似事件，特别是适应性免疫在感染后期的功能。
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