European Laboratory for the Investigation of Food Induced Diseases, Department of Translational Medical Science, Section of Pediatrics, and ELFID, University Federico II, Via S. Pansini 5, 80131 Naples, Italy.
Department of Gynecology, Leiden University Medical Centre, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
Int J Mol Sci. 2021 Mar 8;22(5):2708. doi: 10.3390/ijms22052708.
Celiac disease (CD) is a frequent intestinal inflammatory disease occurring in genetically susceptible individuals upon gluten ingestion. Recent studies point to a role in CD for genes involved in cell shape, adhesion and actin rearrangements, including a Rho family regulator, Rho GTPase-activating protein 31 (ARHGAP31). In this study, we investigated the morphology and actin cytoskeletons of peripheral monocyte-derived dendritic cells (DCs) from children with CD and controls when in contact with a physiological substrate, fibronectin. DCs were generated from peripheral blood monocytes of pediatric CD patients and controls. After adhesion on fibronectin, DCs showed a higher number of protrusions and a more elongated shape in CD patients compared with controls, as assessed by immunofluorescence actin staining, transmitted light staining and video time-lapse microscopy. These alterations did not depend on active intestinal inflammation associated with gluten consumption and were specific to CD, since they were not found in subjects affected by other intestinal inflammatory conditions. The elongated morphology was not a result of differences in DC activation or maturation status, and did not depend on the human leukocyte antigen (HLA)-DQ2 haplotype. Notably, we found that ARH-GAP31 mRNA levels were decreased while RhoA-GTP activity was increased in CD DCs, pointing to an impairment of the Rho pathway in CD cells. Accordingly, Rho inhibition was able to prevent the cytoskeleton rearrangements leading to the elongated morphology of celiac DCs upon adhesion on fibronectin, confirming the role of this pathway in the observed phenotype. In conclusion, adhesion on fibronectin discriminated CD from the controls' DCs, revealing a gluten-independent CD-specific cellular phenotype related to DC shape and regulated by RhoA activity.
乳糜泻(CD)是一种常见的肠道炎症性疾病,发生在对麸质摄入具有遗传易感性的个体中。最近的研究表明,细胞形状、黏附和肌动蛋白重排相关基因,包括 Rho 家族调节剂 Rho GTPase 激活蛋白 31(ARHGAP31),在 CD 中起作用。在这项研究中,我们研究了与生理底物纤维连接蛋白接触时,来自 CD 患儿和对照者的外周单核细胞衍生树突状细胞(DC)的形态和肌动蛋白细胞骨架。从儿科 CD 患者和对照者的外周血单核细胞中生成 DC。与纤维连接蛋白黏附后,通过免疫荧光肌动蛋白染色、透射光染色和视频延时显微镜评估,与对照者相比,CD 患者的 DC 表现出更多的突起和更长的形状。这些改变不依赖于与麸质摄入相关的活跃肠道炎症,并且是 CD 的特异性改变,因为它们在患有其他肠道炎症性疾病的患者中未发现。这种拉长的形态不是由于 DC 激活或成熟状态的差异引起的,也不依赖于人类白细胞抗原(HLA)-DQ2 单倍型。值得注意的是,我们发现 CD DCs 中的 ARH-GAP31 mRNA 水平降低,而 RhoA-GTP 活性增加,表明 CD 细胞中的 Rho 途径受损。因此,Rho 抑制能够防止细胞骨架重排,从而防止 CD DCs 在黏附于纤维连接蛋白时出现拉长形态,证实了该途径在观察到的表型中的作用。总之,纤维连接蛋白黏附可将 CD 与对照者的 DCs 区分开来,揭示了一种与 DC 形状相关且受 RhoA 活性调节的与麸质无关的 CD 特异性细胞表型。