PMID- 33800150 OWN - NLM STAT- MEDLINE DCOM- 20210422 LR - 20210422 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 22 IP - 5 DP - 2021 Mar 8 TI - Pediatric Celiac Disease Patients Show Alterations of Dendritic Cell Shape and Actin Rearrangement. LID - 10.3390/ijms22052708 [doi] LID - 2708 AB - 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. FAU - Discepolo, Valentina AU - Discepolo V AD - 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. FAU - Lania, Giuliana AU - Lania G AD - 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. FAU - Ten Eikelder, Maria Leonarda Gertrude AU - Ten Eikelder MLG AD - Department of Gynecology, Leiden University Medical Centre, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. FAU - Nanayakkara, Merlin AU - Nanayakkara M AD - 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. FAU - Sepe, Leandra AU - Sepe L AD - Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Via S. Pansini 5, 80131 Naples, Italy. FAU - Tufano, Rossella AU - Tufano R AD - Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Via S. Pansini 5, 80131 Naples, Italy. FAU - Troncone, Riccardo AU - Troncone R AD - 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. FAU - Auricchio, Salvatore AU - Auricchio S AD - 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. FAU - Auricchio, Renata AU - Auricchio R AD - 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. FAU - Paolella, Giovanni AU - Paolella G AD - 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. FAU - Barone, Maria Vittoria AU - Barone MV AUID- ORCID: 0000-0001-6190-4917 AD - 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. LA - eng PT - Clinical Trial PT - Journal Article DEP - 20210308 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (ARHGAP31 protein, human) RN - 0 (Actins) RN - 0 (Fibronectins) RN - 0 (GTPase-Activating Proteins) RN - 0 (HLA-DQ Antigens) RN - 0 (HLA-DQ2 antigen) RN - 0 (Phosphoproteins) RN - 124671-05-2 (RHOA protein, human) RN - EC 3.6.5.2 (rhoA GTP-Binding Protein) SB - IM MH - Actins/*metabolism MH - Celiac Disease/*metabolism/pathology MH - Cell Adhesion MH - *Cell Shape MH - Child MH - Child, Preschool MH - Dendritic Cells/*immunology/pathology MH - Female MH - Fibronectins/metabolism MH - GTPase-Activating Proteins/metabolism MH - HLA-DQ Antigens/metabolism MH - Humans MH - Male MH - Monocytes/*metabolism/pathology MH - Phosphoproteins/metabolism MH - rhoA GTP-Binding Protein/metabolism PMC - PMC7962447 OTO - NOTNLM OT - ARHGAP31 OT - RhoA OT - actin cytoskeleton OT - adhesion OT - celiac disease OT - cell shape OT - dendritic cells OT - fibronectin COIS- The authors declare no conflict of interest. EDAT- 2021/04/04 06:00 MHDA- 2021/04/23 06:00 PMCR- 2021/03/08 CRDT- 2021/04/03 01:04 PHST- 2021/02/03 00:00 [received] PHST- 2021/02/24 00:00 [revised] PHST- 2021/02/24 00:00 [accepted] PHST- 2021/04/03 01:04 [entrez] PHST- 2021/04/04 06:00 [pubmed] PHST- 2021/04/23 06:00 [medline] PHST- 2021/03/08 00:00 [pmc-release] AID - ijms22052708 [pii] AID - ijms-22-02708 [pii] AID - 10.3390/ijms22052708 [doi] PST - epublish SO - Int J Mol Sci. 2021 Mar 8;22(5):2708. doi: 10.3390/ijms22052708.