The gut microbiota of toll-like receptor 2-deficient mice exhibits lineage-specific modifications
The gut microbiota of toll-like receptor 2-deficient mice exhibits lineage-specific modifications
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DOI：
10.1111/j.1758-2229.2008.00006.x
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发表时间：
2009-02-01
影响因子：
3.3
通讯作者：
Gophna, Uri
中科院分区：
文献类型：
--
作者：
Albert, Eric J.;Sommerfeld, Katrin;Gophna, Uri
介导哺乳动物先天性免疫系统识别细菌的Toll样受体的突变有可能极大地改变个体微生物区系的组成。在这里，我们通过比较Toll样受体2缺陷小鼠的肠道微生物区系来验证这一假设，无论是年轻的还是中年的小鼠，与具有相同遗传背景的野生型小鼠的肠道微生物区系进行了比较，这些小鼠都生活在特定的无病原体条件下。从小鼠盲肠组织中提取细菌DNA，用细菌16S核糖体RNA基因通用引物扩增，克隆到载体上。选择含有插入片段的克隆进行高通量测序，并对序列数据进行分析，产生物种水平的系统发育数据。用UniFrac软件对克隆文库进行系统发育分析。虽然突变小鼠和野生型小鼠在系统发育种群结构上的配对差异没有统计学意义，但ANOSIM分析确实证明了Toll样受体2缺陷小鼠和它们的野生型小鼠之间的显著差异。观察到的差异可能是由于两种不同的幽门螺杆菌系统型在Toll样受体2缺陷小鼠中高度定植，而野生型小鼠完全不存在。主坐标分析聚类分析表明，在小鼠盲肠微生物区系中，年龄是比基因和母体遗传更弱的决定因素。这些发现表明，尽管Toll样受体的突变可能会导致对特定机会性细菌的易感性增加，但它们并不会显著改变微生物区系的系统发育结构。
Mutations in toll-like receptors that mediate bacterial recognition by the mammalian innate immune system have the potential to substantially alter the composition of an individual's microbiota. Here we tested this hypothesis by comparing the intestinal microbiota of toll-like receptor 2-deficient mice, both young and middle aged, with that of wild-type mice of the same genetic background, housed together under specific pathogen-free conditions. Bacterial DNA was extracted from mouse caecal tissue samples, amplified using universal bacterial 16S ribosomal RNA gene primers, and cloned into a plasmid vector. Insert-containing colonies were picked for high-throughput sequencing, and sequence data were analysed yielding species-level phylogenetic data. Clone libraries were compared by phylogenetic composition analysis using UniFrac. While pairwise differences in phylogenetic population structure between mutant and wildtype mice were not statistically significant, ANOSIM analysis did demonstrate a significant difference between toll-like receptor 2-deficient mice and their wild-type counterparts. The difference observed was probably due to a high level of colonization of the toll-like receptor 2-deficient mice by two distinct Helicobacter phylotypes that were totally absent from wild-type mice. Principal coordinate analysis clustering indicated that age is a weaker determinate than genotype and maternal heritage in the mouse caecal microbiota. The findings suggest that although mutations in toll-like receptors may cause increased susceptibility to specific opportunistic bacteria, they do not dramatically alter the phylogenetic structure of microbiota.