title: Regulatory T cell levels and cytokine production in active non-infectious uveitis: in-vitro effects of pharmacological treatment.
abstract: The aim of this study was to quantify the proportion of regulatory T cells (Treg ) and cytokine expression by peripheral blood mononuclear cells (PBMCs) in patients with active non-infectious uveitis, and to evaluate the effect of in-vitro treatment with infliximab, dexamethasone and cyclosporin A on Treg levels and cytokine production in PBMCs from uveitis patients and healthy subjects. We included a group of 21 patients with active non-infectious uveitis and 18 age-matched healthy subjects. The proportion of forkhead box protein 3 (FoxP3)(+) Treg cells and intracellular tumour necrosis factor (TNF)-α expression in CD4(+) T cells was determined by flow cytometry. PBMCs were also either rested or activated with anti-CD3/anti-CD28 and cultured in the presence or absence of dexamethasone, cyclosporin A and infliximab. Supernatants of cultured PBMCs were collected and TNF-α, interleukin (IL)-10, IL-17 and interferon (IFN)-γ levels were measured by enzyme-linked immunosorbent assay (ELISA). No significant differences were observed in nTreg levels between uveitis patients and healthy subjects. However, PBMCs from uveitis patients produced significantly higher amounts of TNF-α and lower amounts of IL-10. Dexamethasone treatment in vitro significantly reduced FoxP3(+) Treg levels in PBMCs from both healthy subjects and uveitis patients, and all tested drugs significantly reduced TNF-α production in PBMCs. Dexamethasone and cyclosporin A significantly reduced IL-17 and IFN-γ production in PBMCs and dexamethasone up-regulated IL-10 production in activated PBMCs from healthy subjects. Our results suggest that PBMCs from patients with uveitis express more TNF-α and less IL-10 than healthy subjects, and this is independent of FoxP3(+) Treg levels. Treatment with infliximab, dexamethasone and cyclosporin A in vitro modulates cytokine production, but does not increase the proportion of FoxP3(+) Treg cells.
authors.affiliation: Institut Clinic d'Oftalmologia, Hospital Clinic de Barcelona, University of Barcelona, Barcelona, Spain; Fundacio Clinic per la Recerca Biomedica, IDIBAPS, Barcelona, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Molins
authors.forename: M
authors.initials: M
authors.lastname: Mesquida
authors.forename: R W J
authors.initials: RW
authors.lastname: Lee
authors.forename: V
authors.initials: V
authors.lastname: Llorenç
authors.forename: L
authors.initials: L
authors.lastname: Pelegrín
authors.forename: A
authors.initials: A
authors.lastname: Adán
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology*; D018450:Disease Progression; D005260:Female; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D000069285:Infliximab; D018655:Lymphocyte Count; D008297:Male; D008875:Middle Aged; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000276:immunology; D014605:Uveitis / Q000188:drug therapy* / Q000276:immunology; D055815:Young Adult
chemical_list: D000911:Antibodies, Monoclonal; D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D003907:Dexamethasone; D016572:Cyclosporine; D000069285:Infliximab
keywords: IL-10; TNF-α; regulatory T cell; uveitis