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Characteristics of Gut Microbiota and Their Association with Lymphocyte Subsets and Disease Activity in Ankylosing Spondylitis
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摘要:
目的 探究强直性脊柱炎(ankylosing spondylitis, AS)患者肠道菌群变化特征及其与疾病活动度、外周血淋巴细胞亚群的关系。 方法 本研究为回顾性分析。研究对象为2019年12月至2020年6月山西医科大学第二医院住院治疗的AS患者及性别、年龄与之相匹配的健康人群。收集两组肠道菌群生物信息学分析结果以及AS患者外周血淋巴细胞亚群和疾病活动度指标。比较两组肠道菌群差异，并采用Spearman相关法分析AS患者肠道菌群与疾病活动度、外周血淋巴细胞亚群的相关性。 结果 共入选符合纳入和排除标准的AS患者62例(低、高、极高疾病活动度分别为11例、26例、25例)，健康人群62名。α多样性分析显示，AS患者肠道菌群Chao1指数和ACE指数均低于健康人群(P均＜0.05)；β多样性分析显示，两组菌群结构存在差异(P＜0.01)。在肠道微生态构成分析中，发现两组样本肠道菌群优势菌门均以厚壁菌门、拟杆菌门、变形菌门为主，但二者在门和属水平上多种菌群的相对丰度存在差异。在Stamp差异菌群分析中，AS患者在门、属水平亦显示出不同于健康人群的特征：在门水平上，AS患者变形菌门、髌骨细菌门等菌群的相对丰度升高(P均＜0.05)，厚壁菌门、梭杆菌门等菌群的相对丰度降低(P均＜0.05)；在属水平上，AS患者大肠杆菌志贺菌属、克雷伯氏菌属、肠球菌属等菌群的相对丰度升高(P均＜0.05)，普雷沃氏菌属、粪杆菌属等菌群的相对丰度降低(P均＜0.05)。Spearman相关性分析表明，AS患者粪杆菌属、瘤胃球菌属及克雷伯氏菌属等菌属的相对丰度与疾病活动度或其相关指标呈正相关(P均＜0.05)；阿加杆菌属的相对丰度与T细胞(r=0.302，P=0.017)、CD4+T细胞(r=0.310，P=0.014)、B细胞(r=0.292，P=0.021)、Th2细胞(r=0.429，P＜0.001)、Th17细胞(r=0.288，P=0.023)水平，链球菌属的相对丰度与B细胞水平(r=0.270，P=0.034)，普雷沃氏菌属的相对丰度与Th1细胞(r=0.279，P=0.028)、Th17细胞(r=0.262，P=0.040)水平，CAG-352菌属的相对丰度与Th1细胞水平(r=0.283，P=0.030)均呈正相关；大肠杆菌志贺菌属的相对丰度与Th2细胞水平(r=-0.261，P=0.040)，其他肠杆菌科细菌属的相对丰度与CD4+T细胞水平(r=-0.255，P=0.046)均呈负相关。 结论 AS患者肠道菌群多样性降低，致病菌表达增多，且与外周血淋巴细胞亚群和疾病活动度具有相关性，可能参与了AS的发生与发展。 Abstract:Objective To investigate the characteristics of gut microbiome and their associations with lymphocyte subsets and disease activity in patients with ankylosing spondylitis (AS). Methods This study was a retrospective analysis. The subjects of the study were AS patients who were hospitalized in the Second Hospital of Shanxi Medical University from December 2019 to June 2020, as well as gender- and age-matched healthy controls (HCs). The fecal samples were collected, and the V3-V4 variable regions of 16S rRNA gene of gut microbiome were sequenced for bioinformatics analysis. Peripheral venous blood was collected from AS patients to determine peripheral blood lymphocyte subsets and disease activity indicators. Spearman correlation test was used to analyze the correlations between the relative abundances of gut microbiota and peripheral blood lymphocyte subsets as well as disease activity in AS patients. Results A total of 62 AS patients (11 with low disease activity, 26 with high disease activity, and 25 with extremely high disease activity) and 62 healthy people who met the inclusion and exclusion criteria were enrolled. As for α-diversity, ACE and Chao1 indices were lower in AS than in HCs(P < 0.05). Bray curtis distance-based β-diversity analysis revealed significant difference in the microbial community between AS and HCs (P < 0.01). As for the composition of the gut microbiome, Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla in the gut microbiota of both groups, but there were differences in the abundance of various bacteria at the phylum and genus levels. In Stamp analysis, fecal microbial communities in AS differed significantly from those in HCs, which were characterized by higher abundances of phylum Proteobacteria and Patescibacteria(all P < 0.05) and a lower abundance of phylum Firmicutes and Fusobacteriota (all P < 0.05). At the genus level, the abundances of Escherichia-Shigella, Klebsiella and Enterococcus were increased while those of Prevotella and Faecalibacterium were decreased in AS patients compared to HCs(all P < 0.05). Spearman correlation analysis showed that the relative abundances of Faecalibacterium, Ruminococcus and Klebsiella in AS patients were significantly positively correlated with disease activity or its related indicators(all P < 0.05). There were positive correlations between Agathobacter and T cell (r=0.302, P=0.017), CD4+T cell (r=0.310, P=0.014), B cell (r=0.292, P=0.021), Th2 cell (r=0.429, P < 0.001), Th17 cell (r=0.288, P=0.023), Streptococcus and B cell (r=0.270, P=0.034), Prevotella and Th1 cell (r=0.279, P=0.028), Th17 cell (r=0.262, P=0.040), CAG-352 and Th1 cell (r=0.283, P=0.030). There were negative correlations between Escherichia-Shigella and Th2 cell(r=-0.261, P=0.040), other Enterobacteriaceae and CD4+T cell (r=-0.255, P=0.046). Conclusions The diversity of gut microbiota is reduced in AS patients. The abundance of pathogenic bacteria in AS patients is increased, which is correlated with changes in peripheral blood lymphocyte subsets and disease activity. Dysbiosis may be involved in the occurrence and development of AS. -
Key words:
- ankylosing spondylitis /
- gut microbiota /
- lymphocyte subsets /
- immune system diseases
作者贡献：宋子怡负责选题设计并撰写论文初稿；张升校、乔军负责实验和数据分析；赵蓉、宋珊、程婷负责数据收集和文献查阅；王彩虹、李小峰负责论文修订。利益冲突：所有作者均声明不存在利益冲突 -
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