title: Interaction among activated lymphocytes and mesenchymal cells through podoplanin is critical for a high IL-17 secretion.
abstract: During chronic inflammation, immune cells, notably Th17 cells, infiltrate the inflammatory site and interact with local mesenchymal cells. Applied to rheumatoid arthritis (RA), the aim is to study the interactions between synoviocytes and peripheral blood mononuclear cells (PBMC) with a focus on the Th17 pathway and to identify a mechanism which leads to high IL-17 secretion with an interest on podoplanin.
  
  
  
  PBMC from healthy donors and RA patients were co-cultured with RA synoviocytes during 48 h, in the presence or not of phytohemagglutinin. An antibody against podoplanin was used in co-culture. Cytokine production (IL-6, IL-1β and IL-17) was measured by ELISA and cell staining (CD3, CD4, IL-17 and podoplanin) by flow cytometry.
  
  
  
  In control conditions, IL-6 and IL-1β production was increased in PBMC-synoviocyte co-culture compared to PBMC alone (p = 0.02). No additional effect was observed with PBMC activation. Flow cytometry analysis showed no difference in the percentage of Th17 cells in activated PBMC alone or with synoviocytes (p = 0.4), indicating that Th17 differentiation requires only T cell activation. Conversely, IL-17 production was highly increased in co-cultures with activated PBMC vs. activated PBMC alone (p = 0.002). Transwell experiments confirm that cell-cell contact was critical for IL-17 secretion. The incubation of either PBMC or synoviocytes with an anti-podoplanin antibody decreased IL-17 secretion by 60 % (p = 0.008).
  
  
  
  Interactions between resting PBMC and synoviocytes are sufficient to induce IL-6 and IL-1β production. Both PBMC activation and cell interactions are needed to induce a high IL-17 secretion. Podoplanin contributes at the level of both lymphocytes and synoviocytes.
authors.affiliation: Immunogenomics and Inflammation Research Unit, EA 4130, Edouard Herriot Hospital, Hospices Civils de Lyon and University Claude Bernard Lyon 1, Place d'Arsonval, Lyon, 69003, France.
authors.forename: Mélissa
authors.initials: M
authors.lastname: Noack
authors.affiliation: Immunogenomics and Inflammation Research Unit, EA 4130, Edouard Herriot Hospital, Hospices Civils de Lyon and University Claude Bernard Lyon 1, Place d'Arsonval, Lyon, 69003, France.
authors.forename: Ndiémé
authors.initials: N
authors.lastname: Ndongo-Thiam
authors.affiliation: Immunogenomics and Inflammation Research Unit, EA 4130, Edouard Herriot Hospital, Hospices Civils de Lyon and University Claude Bernard Lyon 1, Place d'Arsonval, Lyon, 69003, France. miossec@univ-lyon1.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Miossec
mesh_terms: D001172:Arthritis, Rheumatoid / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D018920:Coculture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D006801:Humans; D020381:Interleukin-17 / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology*; D060888:Real-Time Polymerase Chain Reaction; D000070918:Synoviocytes / Q000276:immunology*; D058504:Th17 Cells / Q000276:immunology* / Q000378:metabolism; D000098853:Podoplanin
chemical_list: D020381:Interleukin-17; D008562:Membrane Glycoproteins; C508717:IL17A protein, human; C117468:PDPN protein, human; D000098853:Podoplanin
keywords: Cell interactions; IL-17; Podoplanin; Rheumatoid arthritis; Synoviocytes