Journal Article

Immunomodulatory Effects of Bone Marrow-Derived Mesenchymal Stem Cells on Pro-Inflammatory Cytokine-Stimulated Human Corneal Epithelial Cells

McClellan, Kathryn, Sutton, Gerard, King, Nicholas J. C., Petsoglou, Con, Wen, Li, Zhu, Meidong, Madigan, Michele C., You, Jingjing, Billson, Francis A.

2014

Overview

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. 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. 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. 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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adhesion / Analysis / Animals / Anti-inflammatory agents / Biological response modifiers / Biology and Life Sciences / Bone marrow / Bone morphogenetic proteins / Cell adhesion & migration / Cell Adhesion Molecules - metabolism / Cell culture / Cell cycle / Cell Line, Transformed / Cell surface / Conditioning / Cornea / Cytokines / Cytokines - pharmacology / Cytometry / Diabetes / Enzyme Activation / Enzyme-linked immunosorbent assay / Enzymes / Epithelial cells / Epithelial Cells - drug effects / Epithelial Cells - immunology / Epithelial Cells - metabolism / Epithelium / Epithelium, Corneal - cytology / Epithelium, Corneal - drug effects / Epithelium, Corneal - immunology / Epithelium, Corneal - metabolism / Flow cytometry / Gene expression / Growth factors / Histocompatibility antigen HLA / Histocompatibility antigens / HLA histocompatibility antigens / Humans / Immunofluorescence / Immunomodulation / Immunophenotyping / Immunoregulation / In vitro methods and tests / Indoleamine-Pyrrole 2,3,-Dioxygenase - metabolism / Inflammation / Inflammation Mediators - pharmacology / Intercellular adhesion molecule 1 / Interferon / Interferon gamma / Kinases / Leukocytes / Localization / Medicine and Health Sciences / Mesenchymal stem cells / Mesenchymal Stromal Cells - immunology / Mesenchymal Stromal Cells - metabolism / Mesenchyme / Monoculture / NF-kappa B - metabolism / Phenotype / Protein Transport / Rats / Rodents / Signal transduction / Signaling / Stem cells / Transcription / Transforming Growth Factor beta1 - metabolism / Transforming growth factor-b1 / Transforming growth factors / Transplants & implants / Tryptophan 2,3-dioxygenase / Tumor necrosis factor / Tumor necrosis factor-TNF / Wound healing / γ-Interferon