摘要
目的解析强直性脊柱炎(ankylosing spondylitis,AS)患者合并克罗恩病(Crohn′s disease,CD)的肠黏膜及外周血炎症微环境中免疫细胞浸润特征、炎症相关信号通路的活化情况。方法对基因表达综合数据库(Gene Expression Omnibus,GEO)中包含CD的AS患者肠黏膜芯片数据进行CIBERSORT反卷积免疫细胞浸润及基因集富集分析(gene set enrichment analysis,GSEA),探索炎症免疫细胞浸润及通路活化情况,同时对AS合并CD的肠黏膜及外周血的单细胞测序数据集进行细胞亚群T细胞亚群比例比较以及进行炎症相关信号通路的活化情况验证。结果利用CIBERSORT算法计算芯片样本中不同免疫细胞亚型的百分比,发现AS肠道黏膜组织中CD8^(+)T淋巴细胞的含量随着疾病的发展而增加,从无肠炎到亚临床肠炎再到肠炎时,CD8^(+)T淋巴细胞的含量逐渐增加。对肠黏膜及外周血的淋巴细胞进行单细胞亚群提取并进行炎症相关通路基因集活性打分,发现CD8^(+)T淋巴细胞在肠黏膜中TNFA-SIGNALING-VIA-NFKB、INTERFERON-GAMMA-RESPONSE、INFLAMMATORY-RESPONSE在AS合并CD分组中较AS组的通路活性明显升高,但IL17-SIGNALING-PATHWAY无明显激活;而在外周血中,CD8^(+)T淋巴细胞的TNFA-SIGNALING-VIA-NFKB、INTERFERON-GAMMA-RESPONSE、INFLAMMATORY-RESPONSE、IL17-SIGNALING-PATHWAY的通路活性则表现为AS合并CD组比AS组明显升高。单细胞测序中CD8^(+)T细胞在AS合并CD患者外周血中比例增高。结论AS合并CD患者肠黏膜中TNF-α、IFN-γ相关的炎症信号通路明显激活,外周血CD8^(+)T淋巴细胞比例增高,可作为判断AS合并肠炎的参考指标。
Objective To analyze the characteristics of immune cell infiltration and activation of inflammation-related signaling pathways in the intestinal mucosa and peripheral blood inflammatory microenvironment of patients with ankylosing spondylitis(AS)coexisting with Crohn′s Disease(CD).Methods CIBERSORT deconvolution of immune cell infiltration and gene set enrichment analysis(GSEA)were performed on microarray data from AS patients with CD obtained from the Gene Expression Omnibus(GEO)database.This analysis explored the infiltration of inflammatory immune cells and the activation status of inflammatory-related signaling pathways.Additionally,single-cell sequencing data from the intestinal mucosa and peripheral blood of AS patients with CD were used to compare the proportions of T cell subgroups and validate the activation status of inflammation-related signaling pathways.Results Using the CIBERSORT algorithm to calculate the percentage of different immune cell subtypes in the chip samples,it was found that the content of CD8^(+)T lymphocytes in the intestinal mucosal tissue of AS patients increased gradually with the progression of the disease,from non-enteritis to subclinical enteritis,and then to enteritis.Single-cell subgroup extraction and scoring for the activation of inflammation-related pathway gene sets in lymphocytes from the intestinal mucosa and peripheral blood revealed that CD8^(+)T lymphocytes in the intestinal mucosa of AS with CD had significantly increased activity in the TNFA-SIGNALING-VIA-NFKB,INTERFERON-GAMMA-RESPONSE,and INFLAMMATORY-RESPONSE pathways compared to AS alone,but IL17-SIGNALING-PATHWAY showed no significant activation.In peripheral blood,the pathway activities of TNFA-SIGNALING-VIA-NFKB,INTERFERON-GAMMA-RESPONSE,INFLAMMATORY-RESPONSE,and IL17-SIGNALING-PATHWAY in CD8^(+)T lymphocytes were significantly higher in the AS with CD group compared to the AS group.Single-cell sequencing also revealed an increased proportion of CD8^(+)T cells in the peripheral blood of AS patients with CD.Conclusion In AS patients with CD,the inflammatory signaling pathways related to TNF-αand IFN-γare significantly activated in the intestinal mucosa.The proportion of CD8^(+)T lymphocytes in peripheral blood is increased,serving as a potential indicator for determining the coexistence of AS and enteritis.
作者
罗锡庆
古洁若
LUO Xi-qing;GU Jie-ruo(Guangdong Provincial Clinical Medical Research Center of Immune Diseases,Department of Rheumatology and Immunology,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,Guangdong,China;不详)
出处
《广东医学》
CAS
2024年第1期50-62,共13页
Guangdong Medical Journal
基金
广东省重点领域研发计划项目(2023B1111030002)
广东省免疫疾病临床医学研究中心(2020B1111170008)。
关键词
强直性脊柱炎
单细胞测序
炎症微环境
ankylosing spondylitis
single cell sequencing
inflammatory microenvironment
