May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Inflammatory Cytokine Effects on Co–Stimulatory Molecule and Chemokine Expression in the Human Conjunctival Epithelial Cell Line, IOBA–NHC
Author Affiliations & Notes
  • K.F. Siemasko
    Biological Sciences, Allergan, Irvine, CA
  • J. Gao
    Biological Sciences, Allergan, Irvine, CA
  • A. Enriquez de Salamanca
    IOBA–University of Valladolid, Valladolid, Spain
  • Y. Diebold
    IOBA–University of Valladolid, Valladolid, Spain
  • V.L. Calder
    University College London, London, United Kingdom
  • M. Calonge
    IOBA–University of Valladolid, Valladolid, Spain
  • M.E. Stern
    Biological Sciences, Allergan, Irvine, CA
  • Footnotes
    Commercial Relationships  K.F. Siemasko, Allergan, Inc. E; J. Gao, Allergan, Inc. E; A. Enriquez de Salamanca, None; Y. Diebold, None; V.L. Calder, Allergan, Inc. C; M. Calonge, Allergan, Inc. C; M.E. Stern, Allergan, Inc. E.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 2687. doi:
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      K.F. Siemasko, J. Gao, A. Enriquez de Salamanca, Y. Diebold, V.L. Calder, M. Calonge, M.E. Stern; Inflammatory Cytokine Effects on Co–Stimulatory Molecule and Chemokine Expression in the Human Conjunctival Epithelial Cell Line, IOBA–NHC . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2687.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Keratoconjunctivitis sicca is mediated by T cell activation that results in ocular surface inflammation. T cell activation requires two signals: 1) MHC–peptide binding to the TCR complex and 2) co–stimulatory molecule activation of cell surface molecules such as CD28 on the T cell binding to B7 molecules on the antigen presenting cell or CD40 ligand binding to CD40. T cells can be recruited to inflammatory sites by the chemokines IL–8 and RANTES. We previously reported that the normal human conjunctival cell line IOBA–NHC express HLA–DR when stimulated with IFN–γ. The purpose of this study was to determine if IOBA–NHC cells, in the presence of inflammatory cytokines, can provide co–stimulatory signals for T cell activation and chemokines required for T cell recruitment to the site of inflammation.Methods:IOBA–NHC were treated with human recombinant IFN–γ (10–1000 U/ml), TNF–α (0.1–100 ng/ml), IL–1ß (0.1–100 ng/ml), or IL–6 (0.1–100 ng/ml) for 72 hrs. Cells were incubated with specific FITC or PE–conjugated co–stimulatory antibodies against B7.1, B7.2, or CD40 and analyzed by flow cytometry. Secretion of the chemokines IL–8 and RANTES were determined by analyzing the supernatants of stimulated cells by ELISA. Results:IOBA–NHC cells do not constitutively express detectable levels of B7.1, B7.2, or CD40 as determined by flow cytometry. In the presence of IFN–γ, IOBA–NHC expressed CD40, but not B7.1 or B7.2. Co–culture of IFN–γ–stimulated IOBA cells with Jurkat T cells induced IOBA–NHC to express B7.1 and B7.2 (Gao et al., 2003 ARVO). These results suggest that the interaction of CD40 on the IOBA–NHC with its ligand on the T cell is required for B7 expression. Upon examination of cytokine induced chemokine expression, IOBA–NHC cells secreted IL–8 in the presence of all TNF–α concentrations tested. TNF–α and/or IFN–γ induced RANTES secretion. IL–1ß and IL–6 had no detectable effect on any of the markers tested. Conclusions: Co–stimulatory molecule and chemokine expression by IOBA–NHC suggests that conjunctival epithelial cells may play an essential role in inflammation. The interaction of activated T cells with resident epithelial cells may be important for pathogenesis of ocular inflammatory disease.

Keywords: conjunctiva • inflammation • cytokines/chemokines 
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