Heterogeneity of CD4+CD25+Foxp3+Treg TCR β CDR3 Repertoire Based on the Differences of Symbiotic Microorganisms in the Gut of Mice
Heterogeneity of CD4+CD25+Foxp3+Treg TCR β CDR3 Repertoire Based on the Differences of Symbiotic Microorganisms in the Gut of Mice
复制标题
基于小鼠肠道共生微生物差异的 CD4 CD25 Foxp3 Treg TCR β CDR3 库的异质性
DOI：
10.3389/fcell.2020.576445
复制
发表时间：
2020-09-01
影响因子：
5.5
通讯作者：
Yao, Xinsheng
中科院分区：
文献类型：
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作者：
Li, Jun;Xue, Huaijuan;Yao, Xinsheng
肠道微生物在自身免疫性疾病的发生和发展中发挥着至关重要的作用。肠道微生物的多样性受生活环境的影响，调节外周免疫器官和局部组织的免疫功能。采用16 S rDNA序列分析方法，对无菌（GF）、无特定病原体（SPF）和清洁（CL）BALB/c小鼠肠道微生物的多样性进行了研究。采用高通量测序技术分析GF、SPF和CL小鼠肠道和脾脏调节性T细胞（Treg）TCR β链CDR 3组成及特性，探讨肠道微生物组成差异对肠道和脾脏CD 4 + CD 25 + Foxp 3 +Treg TCR β CDR 3组成的影响。我们观察到GF、SPF和CL小鼠具有不同的肠道微生物组成，并且微生物的丰度和数量与饲养环境的水平呈正相关。结果表明，GF小鼠的脾脏和小肠结构不完整，且在肠道Treg TCRβ CDR 3基因库中，GF小鼠、SPF小鼠和CL小鼠的高频率独特CDR 3氨基酸序列的使用频率存在显著差异，TRBV、TRBJ和TRBV-TRBJ组合基因片段的使用频率存在较大的异质性。但不同饲养环境对小鼠脾脏Treg TCRβ CDR 3库的影响较弱，提示肠道菌群组成的不同可能主要影响局部Treg TCRβ CDR 3库的多样性，并不改变循环免疫系统的整体特性。这些结果为进一步分析肠道微生物调节肠粘膜免疫系统的机制提供了基础数据。
Gut microbes play a crucial role in the occurrence and development of autoimmune diseases. The diversity of intestinal microorganisms affected by the living environment, regulate the immune function of peripheral immune organs and local tissues. In the study, the diversity of intestinal microorganisms of Germ-free (GF), Specific Pathogen-free (SPF), and Clean (CL) BALB/c mice were conducted by 16S rDNA sequencing. High-throughput sequencing technology was used to analysis the composition and characterization of TCR β chain CDR3 repertoires in Regulatory T cells (Treg) in intestine and spleen of GF, SPF, and CL mice, so as to investigate the effects of differential composition of intestinal microorganisms on the CD4+CD25+Foxp3+Treg TCR β CDR3 repertoire of intestine and spleen. We observed that GF, SPF, and CL mice have different gut microorganism composition, and the abundance and quantity of microorganisms are positively correlated with the level of feeding environment. Interestingly, incomplete structure of spleen and small intestine in GF mice was found. Moreover, a significant difference in the usage of high frequency unique CDR3 amino acid sequences was detected in the intestinal Treg TCRβ CDR3 repertoire among GF, SPF and CL mice, and there were a greater heterogeneity in the usage frequency of TRBV, TRBJ, and TRBV-TRBJ combinations gene segments. However, the effect of different feeding environment on the mice Treg TCRβ CDR3 repertoire of spleen was weak, implying that the different composition of intestinal microbiota may primarily affect the diversity of the local Treg TCRβ CDR3 repertoire and does not alter the overall properties of the circulating immune system. These results provide basic data to further analyze the mechanism of gut microbes regulating the intestinal mucosal immune system.