Barts Liver Centre, Blizard Institute, Queen Mary University of London, London, UK.
Division of Gastroenterology and Hepatology, Medical University of South Carolina, Charleston, SC, USA.
Clin Mol Hepatol. 2023 Apr;29(2):417-432. doi: 10.3350/cmh.2022.0205. Epub 2022 Nov 21.
BACKGROUND/AIMS: Immune and inflammatory cells respond to multiple pathological hits in the development of nonalcoholic steatohepatitis (NASH) and fibrosis. Relatively little is known about how their type and function change through the non-alcoholic fatty liver disease (NAFLD) spectrum. Here we used multi-dimensional mass cytometry and a tailored bioinformatic approach to study circulating immune cells sampled from healthy individuals and people with NAFLD.
Cytometry by time of flight using 36 metal-conjugated antibodies was applied to peripheral blood mononuclear cells (PBMCs) from biopsy-proven NASH fibrosis (late disease), steatosis (early disease), and healthy patients. Supervised and unsupervised analyses were used, findings confirmed, and mechanisms assessed using independent healthy and disease PBMC samples.
Of 36 PBMC clusters, 21 changed between controls and disease samples. Significant differences were observed between diseases stages with changes in T cells and myeloid cells throughout disease and B cell changes in late stages. Semi-supervised gating and re-clustering showed that disease stages were associated with fewer monocytes with active signalling and more inactive NK cells; B and T cells bearing activation markers were reduced in late stages, while B cells bearing co-stimulatory molecules were increased. Functionally, disease states were associated with fewer activated mucosal-associated invariant T cells and reduced toll-like receptor-mediated cytokine production in late disease.
A range of innate and adaptive immune changes begin early in NAFLD, and disease stages are associated with a functionally less active phenotype compared to controls. Further study of the immune response in NAFLD spectrum may give insight into mechanisms of disease with potential clinical application.
背景/目的：免疫和炎症细胞对非酒精性脂肪性肝炎（NASH）和纤维化发展过程中的多种病理刺激作出反应。相对而言，人们对其类型和功能如何通过非酒精性脂肪性肝病（NAFLD）谱发生变化知之甚少。在这里，我们使用多维液质联用技术和定制的生物信息学方法来研究从健康个体和 NAFLD 患者中采集的循环免疫细胞。
使用 36 种金属偶联抗体的飞行时间细胞术应用于活检证实的 NASH 纤维化（晚期疾病）、脂肪变性（早期疾病）和健康患者的外周血单核细胞（PBMC）。采用有监督和无监督分析，使用独立的健康和疾病 PBMC 样本进行验证和机制评估。
在 36 个 PBMC 簇中，有 21 个在对照和疾病样本之间发生变化。在疾病阶段之间观察到显著差异，T 细胞和髓样细胞在整个疾病过程中以及 B 细胞在晚期发生变化。半监督门控和重新聚类表明，疾病阶段与具有活跃信号的单核细胞减少和更多无活性 NK 细胞有关；晚期阶段 B 和 T 细胞携带的激活标志物减少，而携带共刺激分子的 B 细胞增加。功能上，疾病状态与较少的激活黏膜相关不变 T 细胞和晚期疾病中 Toll 样受体介导的细胞因子产生减少有关。
NAFLD 早期就出现了一系列固有和适应性免疫变化，与对照组相比，疾病阶段与功能上不那么活跃的表型相关。进一步研究 NAFLD 谱中的免疫反应可能有助于深入了解疾病机制，并具有潜在的临床应用价值。
