May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Flow Cytometric Comparison of the Expression of Cell Surface Markers on Monocyte Derived Dendritic Cells From Normal and Ocular Allergy Donors
Author Affiliations & Notes
  • B. Manzouri
    Department of Ocular Immunology, Institute of Ophthalmology & Moorfields Eye Hospital, London, United Kingdom
  • D. Fattah
    Department of Asthma Biology, GlaxoSmithKline Medicines Research Centre, Stevenage, United Kingdom
  • M. Ohbayashi
    Department of Ocular Immunology, Institute of Ophthalmology, University College London, London, United Kingdom
  • A. Leonardi
    Department of Neuroscience, Ophthalmology Unit, University of Padua, Padua, Italy
  • F. Larkin
    Department of Ocular Immunology, Institute of Ophthalmology & Moorfields Eye Hospital, London, United Kingdom
  • S.J. Ono
    Department of Ocular Immunology, Institute of Ophthalmology, University College London, London, United Kingdom
  • Footnotes
    Commercial Relationships  B. Manzouri, None; D. Fattah, None; M. Ohbayashi, None; A. Leonardi, None; F. Larkin, None; S.J. Ono, None.
  • Footnotes
    Support  Medical Research Council
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 4971. doi:
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      B. Manzouri, D. Fattah, M. Ohbayashi, A. Leonardi, F. Larkin, S.J. Ono; Flow Cytometric Comparison of the Expression of Cell Surface Markers on Monocyte Derived Dendritic Cells From Normal and Ocular Allergy Donors . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4971.

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

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Abstract

Purpose: : The aim of this study was to compare the differences in the phenotype of cell surface markers on monocyte derived dendritic cells (MoDC) cultured from patients with vernal keratoconjunctivitis (VKC) and healthy control subjects.

Methods: : Peripheral blood was obtained from consented VKC patients (n=12) and healthy subjects (n=8). Positive selection of CD14+ cells was performed from the peripheral blood mononuclear fraction derived after density gradient centrifugation. The monocytes were cultured in DC medium supplemented with interleukin 4 (IL–4) and granulocyte–macrophage colony–stimulating (GM–CSF) factor for 7 days to produce MoDC. These cells were singly stained with different conjugated monoclonal antibodies to a variety of cell surface markers and the expression quantified using flow cytometry.

Results: : MoDCs that were successfully cultured expressed high levels of the human dendritic cell surface marker CD11c. Statistically significant phenotypic differences were seen in the MoDC derived from VKC patients for the co–stimulatory molecules CD86 and CD83 and the adhesion molecule CD58. These were all upregulated in the MoDC from VKC patients. A statistically significant difference was also observed for the adhesion molecule CD11b and the low affinity receptor for IgE (CD23) with these both being downregulated in the MoDC from VKC patients. No difference was observed for the expression of the high affinity IgE receptor (FceRI).

Conclusions: : These results confirm that the MoDC from VKC patients are in a more mature state compared to normal control subjects. This might be due to an altered sensitivity of the atopic monocytes to the cytokine IL–4 and/or GM–CSF. Alternatively, MoDC from atopes may be more potent stimulators of the T–cells as reflected in the upregulation of their co–stimulatory molecules. However, systemic priming of the monocytes cannot be excluded. Further research to look at the phenotype of in vivo dendritic cells from these patients would hence be required.

Keywords: antigen presentation/processing • flow cytometry • conjunctivitis 
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