April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Upregulation of Foxp3 Expression by Recombinant Interferon-alpha Therapy in Behcet’s Disease
Author Affiliations & Notes
  • D. S. Yang
    Ocular Biology & Therapeutics, Institute of Ophthalmology, London, United Kingdom
  • G. Galatowicz
    Ocular Biology & Therapeutics, Institute of Ophthalmology, London, United Kingdom
  • V. L. Calder
    Ocular Biology & Therapeutics, Institute of Ophthalmology, London, United Kingdom
  • S. Lightman
    Ocular Biology & Therapeutics, Institute of Ophthalmology, London, United Kingdom
  • Footnotes
    Commercial Relationships  D.S. Yang, None; G. Galatowicz, None; V.L. Calder, None; S. Lightman, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1535. doi:
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      D. S. Yang, G. Galatowicz, V. L. Calder, S. Lightman; Upregulation of Foxp3 Expression by Recombinant Interferon-alpha Therapy in Behcet’s Disease. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1535.

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

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Abstract

Purpose: : Behcet’s disease (BD) is a multisystem inflammatory disorder. Despite vigorous treatment with immunosuppressive agents, the development of recurrent, sight-threatening ocular disease is still unpredictable. Recently, biological agents including interferon-alpha (IFN-) have revolutionised the treatment and improved the long-term outcome. The effect of IFN- is highly pleiotropic and includes immunomodulatory properties. The aim of this study was to investigate whether IFN- therapy is effective through upregulating the Foxp3-expressing regulatory T cell (Treg) population.

Methods: : Whole blood samples were obtained from BD patients on immunosuppressants with or without 6 months of weekly subcutaneous IFN-2b therapy (Schering-Plough, dose adjusted according to a scale in proportion to body weight). Cells were either unstimulated, stimulated with anti-CD3 [5µg/ml] and anti-CD28 Abs [10µg/ml], or stimulated with PMA [25µg/ml] and ionomycin [10µg/ml] for 24 hours before staining for CD4 and CD25, and intracellularly with Foxp3. After washing, cells were acquired on a FACSCalibur (BD Biosciences) and data analysed using Winlist (Verity Software House).

Results: : The Foxp3 expression within the CD4+CD25high cell population in the untreated cells (n=16) increased from 4.98%±2.44 to 14.29%±3.88 (p<0.02) after 6 months of IFN- therapy. In cells stimulated with anti-CD3/28 Abs (n=16), the level rose from 5.86%±2.41 to 17.27%±4.22 (p=0.005). With PMA/ionomycin stimulation, the Foxp3 expression was also enhanced but was not statistically significant. In contrast, in BD patients on immunosuppressants alone, the Foxp3 expression level only showed a slight increase in the CD4+CD25high cell population from 6.62%±1.98 to 7.92%±2.01 (p=N.S.) for unstimulated cells (n=12). For cells treated with anti-CD3/28 Abs (n=12), the level was unchanged (10.07%±3.54 to 10.98%±2.55, p=N.S.) after 6 months. For cells stimulated with PMA/Ionomycin (n=10), a slight decrease from 27.04%±3.30 to 21.37%±3.75 (p=N.S.) was observed.

Conclusions: : An increase in Foxp3 expression level in the CD4+CD25high cell population was detected in BD patients who received 6-month course of subcutaneous IFN-2b injections.

Keywords: immunomodulation/immunoregulation • autoimmune disease • flow cytometry 
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