Department of Immunology, Faculty of Medicine, Uludag University, Bursa, Turkey.
Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Russia.
PLoS One. 2018 Jun 13;13(6):e0198659. doi: 10.1371/journal.pone.0198659. eCollection 2018.
Brucellosis is a serious infectious disease that continues to be a significant cause of morbidity worldwide and across all ages. Despite early diagnosis and treatment, 10-30% of patients develop chronic brucellosis. Although there have been recent advances in our knowledge of Brucella virulence factors and hosts' immune response to the infection, there is a lack of clear data regarding how the infection bypasses the immune system and becomes chronic. The present study investigated immunological factors and their roles in the transition of brucellosis from an acute to a chronic infection in CD4+ T cells. CD4+ T cells sorted from peripheral blood samples of patients with acute or chronic brucellosis and healthy controls using flow cytometry as well as more than 2000 miRNAs were screened using the GeneSpring GX (Agilent) 13.0 miRNA microarray software and were validated using reverse transcription polymerase chain reaction (RT-qPCR). Compared to acute cases, the expression levels of 28 miRNAs were significantly altered in chronic cases. Apart from one miRNA (miR-4649-3p), 27 miRNAs were not expressed in the acute cases (p <0.05, fold change> 2). According to KEGG pathway analysis, these miRNAs are involved in the regulation of target genes that were previously involved in the MAPK signalling pathway, regulation of the actin cytoskeleton, endocytosis, and protein processing in the endoplasmic reticulum. This indicates the potential role of these miRNAs in the development of chronic brucellosis. We suggest that these miRNAs can be used as markers to determine the transition of the disease into chronicity. This is the first study of miRNA expression that analyses human CD4+ T cells to clarify the mechanism of chronicity in brucellosis.
布鲁氏菌病是一种严重的传染病，仍然是全球和各年龄段发病率的重要原因。尽管早期诊断和治疗，10-30%的患者发展为慢性布鲁氏菌病。尽管最近我们对布鲁氏菌毒力因子和宿主对感染的免疫反应有了更多的了解，但关于感染如何绕过免疫系统并变为慢性的，缺乏明确的数据。本研究调查了免疫因素及其在 CD4+T 细胞中从急性感染向慢性感染转变中的作用。使用流式细胞术从急性或慢性布鲁氏菌病患者和健康对照者的外周血样本中分离 CD4+T 细胞，并使用 GeneSpring GX（安捷伦）13.0 miRNA 微阵列软件筛选超过 2000 个 miRNA，并使用逆转录聚合酶链反应（RT-qPCR）进行验证。与急性病例相比，慢性病例中 28 个 miRNA 的表达水平明显改变。除了一个 miRNA（miR-4649-3p）外，27 个 miRNA 在急性病例中未表达（p<0.05，倍数变化>2）。根据 KEGG 通路分析，这些 miRNA 参与了调节靶基因的作用，这些靶基因先前涉及 MAPK 信号通路、肌动蛋白细胞骨架调节、内吞作用和内质网中的蛋白质加工。这表明这些 miRNA 在慢性布鲁氏菌病的发展中具有潜在作用。我们建议，这些 miRNA 可以用作标志物，以确定疾病向慢性转变。这是第一项分析人类 CD4+T 细胞中 miRNA 表达的研究，以阐明布鲁氏菌病慢性化的机制。
