title: Immunomodulatory effects of bone marrow-derived mesenchymal stem cells on pro-inflammatory cytokine-stimulated human corneal epithelial cells.
abstract: OBJECTIVE
  To investigate the modulatory effect of rat bone marrow mesenchymal stem cells (MSC) on human corneal epithelial cells (HCE-T) stimulated with pro-inflammatory cytokines interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in an in vitro co-cultured model.
  
  
  METHODS
  HCE-T alone and co-cultured with MSC were stimulated with IFN-γ/TNF for 24 and 48 hours or left untreated. The expression of intracellular adhesion molecule (ICAM)-1, human leukocyte antigen ABC, DR and G (HLA-ABC, HLA-DR, HLA-G) were investigated by flow cytometry. Subcellular localization of nuclear factor-kappa B (NF-κB) and expression of indoleamine 2,3-dioxygenase (IDO) were assessed by immunofluorescence staining and western blot. The concentration of transforming growth factor beta 1 (TGF-β1) in the conditioned media from different cultures was evaluated by enzyme-linked immunosorbent assay. NF-κB and TGF-β1 signaling pathway blocking experiments were performed to analyze associations between the expression of cell surface molecules and the NF-κB transcription pathway, and the expression of IDO and TGF-β1 signaling pathway.
  
  
  RESULTS
  IFN-γ/TNF treatment significantly up-regulated expression of ICAM-1, HLA-ABC, and induced de novo expression of HLA-DR and IDO on HCE-T cultured alone, while HLA-G expression remained unaffected. Up-regulation was significantly inhibited by co-culture with MSC. Increased TGF-β1 secretion was detected in 48 h IFN-γ/TNF-stimulated MSC monocultures and HCE-T/MSC co-cultures. MSC attenuated the activation of cytokine-induced NF-κB and IDO induction. Blockade of NF-κB transcription pathway by BMS-345541 significantly reduced the up-regulation of ICAM-1, HLA-ABC, HLA-DR and IDO expression, while blockade of TGF-β1 signaling pathways reversed the modulatory effect of MSC on IDO expression.
  
  
  CONCLUSIONS
  MSC reduced the expression of adhesion and immunoregulatory molecules on pro-inflammatory cytokine-stimulated HCE-T via the NF-κB transcription pathway. MSC attenuated expression of IDO through both NF-κB transcription and TGF-β1 signaling pathways. Co-culture of HCEC with MSC therefore provides a useful in vitro model to study the anti-inflammatory properties of MSC on corneal epithelium.
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Li
authors.initials: L
authors.lastname: Wen
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; Lions New South Wales Eye Bank, NSW Organ and Tissue Donation Service, Sydney, New South Wales, Australia; Sydney Eye Hospital, Sydney, New South Wales, Australia.
authors.forename: Meidong
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; School of Optometry & Vision Science, University of New South Wales, Kensington, New South Wales, Australia.
authors.forename: Michele C
authors.initials: MC
authors.lastname: Madigan
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Jingjing
authors.initials: J
authors.lastname: You
authors.affiliation: Discipline of Pathology, School of Medical Sciences and Bosch Institute, Sydney Medical School, University of Sydney, Camperdown, New South Wales, Australia.
authors.forename: Nicholas J C
authors.initials: NJ
authors.lastname: King
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; Sydney Eye Hospital, Sydney, New South Wales, Australia; Sight for Life Foundation, Sydney, New South Wales, Australia.
authors.forename: Francis A
authors.initials: FA
authors.lastname: Billson
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; Lions New South Wales Eye Bank, NSW Organ and Tissue Donation Service, Sydney, New South Wales, Australia; Sydney Eye Hospital, Sydney, New South Wales, Australia.
authors.forename: Kathryn
authors.initials: K
authors.lastname: McClellan
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; Lions New South Wales Eye Bank, NSW Organ and Tissue Donation Service, Sydney, New South Wales, Australia; Sydney Eye Hospital, Sydney, New South Wales, Australia; Vision Eye Institute, Chatswood, New South Wales, Australia; Auckland University, Auckland, New Zealand.
authors.forename: Gerard
authors.initials: G
authors.lastname: Sutton
authors.affiliation: Save Sight Institute & Discipline of Clinical Ophthalmology, University of Sydney, Sydney, New South Wales, Australia; Lions New South Wales Eye Bank, NSW Organ and Tissue Donation Service, Sydney, New South Wales, Australia; Sydney Eye Hospital, Sydney, New South Wales, Australia.
authors.forename: Con
authors.initials: C
authors.lastname: Petsoglou
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002461:Cell Line, Transformed; D016207:Cytokines / Q000494:pharmacology*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D056747:Immunomodulation; D016130:Immunophenotyping; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000378:metabolism; D018836:Inflammation Mediators / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000276:immunology / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D010641:Phenotype; D021381:Protein Transport; D051381:Rats; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D015815:Cell Adhesion Molecules; D016207:Cytokines; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D018836:Inflammation Mediators; D016328:NF-kappa B; D053773:Transforming Growth Factor beta1