[Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis].

To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject's immune indicators (CD3+, CD4+, CD8+, CD4+CD25+CD127-Treg, CD14+CD16+, CD14+CD16-), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ² test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups (P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group (Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group (P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group (P<0.05). Comparison of immune indexes between groups showed that CD14+CD16+monocytes (%) in the PTB group were higher than those in the HC group (t=2.456, P=0.001 6<0.05), while CD14+CD16-monocytes (%) were lower than HC (t=-4.368, P=0.000<0.05), while the differences in CD3+, CD4+, CD8+, CD4+/CD8+and Treg (CD4+CD25+CD127-) were not statistically significant (P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4+/CD8+, CD14+CD16+(r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14+CD16-Correlation (r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4+/CD8+and CD4+(r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3+, CD8+and CD14+CD16-are positively correlated (r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14+CD16+(r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body's immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.探讨初治肺结核患者肠道菌群改变与机体免疫指标的相关性，为肺结核防治提供参考依据。采取单中心病例对照研究，选取2020年10月至2021年4月于南华大学附属长沙中心医院肺结核科，诊断为初治肺结核的患者43例，同期健康查体的人群43名作为对照组。初治肺结核患者（pulmonary tuberculosis，PTB）43例，同期体检健康者（healthy control，HC）43名，收集受试对象新鲜粪便和全血，采用Illumina Hiseq高通量测序技术对粪便中所有微生物16S rRNA的V4区进行扩增测序，通过QIIME软件分析肠道菌群结构。利用流式细胞术测定受试者免疫指标（CD3+、CD4+、CD8+、CD4+CD25+CD127-Treg、CD14+CD16+、CD14+CD16-），分析初治肺结核患者肠道菌群改变与免疫功能的内在关联。利用χ²检验、t检验、Wilcox秩和检验等方法分析两组人群年龄、性别、α 多样性及菌群相对丰度差异。PTB组与HC组相比，肠道菌群α多样性下降（shannon指数：t=3.906，P=0.000 2；simpson指数：Z=553，P=0.004 7；chao1指数：t=5.395，P= 0.000 0）。β多样性分析显示，两组肠道菌群结构存在明显差异（P=0.000）。物种差异分析显示，在门水平上，PTB组厚壁菌门相对丰度明显低于HC组（Z=486.0，P=0.000 5）。在属水平上，两组间共存在15个差异菌属，PTB组中双歧杆菌属（Bifidobacterium）、肠球菌属（Enterococcus）、乳杆菌属（Lactobacillus）、厌氧棒杆菌属（Anaerostipes）、韦荣球菌属（Veillonella）以上5个菌属相对丰度较HC组高（P<0.05）；丁酸单胞菌（Butyricimonas）、梭状芽胞菌属（Clostridium）、布劳特菌属（Blautia）、粪球菌属（Coprococcus）、多尔菌属（Dorea）、毛螺菌属（Lachnospira）、罗斯菌属（Roseburia）、粪杆菌属（Faecalibacterium）、瘤胃球菌属（Ruminococcus）、小杆菌属（Dialister）等10个菌属相对丰度较HC组低（P<0.05）。组间免疫指标比较结果显示，PTB组CD14+CD16+单核细胞（%）较HC组高（t=2.456，P=0.0016<0.05），而CD14+CD16-单核细胞（%）较HC低（t=-4.368，P=0.000<0.05），而CD3+、CD4+、CD8+、CD4+/CD8+及Treg（CD4+CD25+CD127-）差异无统计学意义（P>0.05）。Spearman相关性分析显示，PTB组中厚壁菌门与CD4+/CD8+、CD14+CD16+呈负相关（r=-0.218，P=0.048；r=-0.245，P=0.025），与CD14+CD16-呈正相关（r=0.250，P=0.022）；在属水平上，粪杆菌属与CD4+/CD8+、CD4+呈正相关（r=0.250，P=0.023；r=0.258，P=0.019）；罗斯菌属与CD3+、CD8+及CD14+CD16-呈正相关（r=0.27，P=0.024；r=0.219，P=0.046；r=0.027，P=0.039），与CD14+CD16+呈负相关（r=-0.280，P=0.01）。初治肺结核患者肠道菌群结构改变，可能是机体免疫功能低下的影响因素之一，有针对性地优化肠道菌群结构，改善机体免疫力，或可作为一种有效辅助治疗肺结核的方式。.