Department of Medical Genetics, Third Faculty of Medicine, Charles University, Ruská 87, 100 00 Prague, Czech Republic.
Department of Immunology and Clinical Biochemistry, Third Faculty of Medicine, Charles University, Ruská 87, 100 00 Prague, Czech Republic.
Int J Mol Sci. 2021 Sep 15;22(18):9982. doi: 10.3390/ijms22189982.
The autoimmune condition, Celiac Disease (CeD), displays broad clinical symptoms due to gluten exposure. Its genetic association with DQ variants in the human leukocyte antigen (HLA) system has been recognised. Monocyte-derived mature dendritic cells (MoDCs) present gluten peptides through HLA-DQ and co-stimulatory molecules to T lymphocytes, eliciting a cytokine-rich microenvironment. Having access to CeD associated families prevalent in the Czech Republic, this study utilised an in vitro model to investigate their differential monocyte profile. The higher monocyte yields isolated from PBMCs of CeD patients versus control individuals also reflected the greater proportion of dendritic cells derived from these sources following lipopolysaccharide (LPS)/ peptic-tryptic-gliadin (PTG) fragment stimulation. Cell surface markers of CeD monocytes and MoDCs were subsequently profiled. This foremost study identified a novel bio-profile characterised by elevated CD64 and reduced CD33 levels, unique to CD14++ monocytes of CeD patients. Normalisation to LPS stimulation revealed the increased sensitivity of CeD-MoDCs to PTG, as shown by CD86 and HLA-DQ flow cytometric readouts. Enhanced CD86 and HLA-DQ expression in CeD-MoDCs were revealed by confocal microscopy. Analysis highlighted their dominance at the CeD-MoDC membrane in comparison to controls, reflective of superior antigen presentation ability. In conclusion, this investigative study deciphered the monocytes and MoDCs of CeD patients with the identification of a novel bio-profile marker of potential diagnostic value for clinical interpretation. Herein, the characterisation of CD86 and HLA-DQ as activators to stimulants, along with robust membrane assembly reflective of efficient antigen presentation, offers CeD targeted therapeutic avenues worth further exploration.
自身免疫性疾病乳糜泻 (CeD) 由于麸质暴露而表现出广泛的临床症状。其与人类白细胞抗原 (HLA) 系统中的 DQ 变体的遗传关联已得到认可。单核细胞衍生的成熟树突状细胞 (MoDC) 通过 HLA-DQ 和共刺激分子将麸质肽呈递给 T 淋巴细胞，引发富含细胞因子的微环境。本研究利用体外模型，研究了捷克共和国常见的乳糜泻相关家族的差异单核细胞谱。从 CeD 患者 PBMC 中分离出的单核细胞产量高于对照个体，这也反映了从这些来源衍生的树突状细胞在脂多糖 (LPS)/ 肽酶-麦醇溶蛋白 (PTG) 片段刺激后比例更高。随后对 CeD 单核细胞和 MoDC 的细胞表面标志物进行了分析。这项首要研究确定了一种新的生物特征，其特征是 CD64 水平升高和 CD33 水平降低，这是 CeD 患者 CD14++单核细胞所特有的。用 LPS 刺激归一化后，发现 CeD-MoDC 对 PTG 的敏感性增加，这表现在 CD86 和 HLA-DQ 流式细胞术读出上。共聚焦显微镜显示 CeD-MoDC 中增强的 CD86 和 HLA-DQ 表达。分析突出了它们在 CeD-MoDC 膜上相对于对照物的优势，反映了优越的抗原呈递能力。总之，这项研究深入分析了 CeD 患者的单核细胞和 MoDC，确定了一种新的生物特征标志物，具有潜在的临床解释诊断价值。在此，CD86 和 HLA-DQ 作为刺激物的激活剂以及反映有效抗原呈递的稳健膜组装的特征，为乳糜泻靶向治疗提供了值得进一步探索的途径。
