May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Major Role of T Cells in the Induction of Il-17+ Uveitogenic T Cells in Mouse Eau
Author Affiliations & Notes
  • C. Lan
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • Y. Peng
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • H. Shao
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • Y. Cui
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • H. J. Kaplan
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • D. Sun
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky
  • Footnotes
    Commercial Relationships  C. Lan, None; Y. Peng, None; H. Shao, None; Y. Cui, None; H.J. Kaplan, None; D. Sun, None.
  • Footnotes
    Support  Supported in part by NIH grants EY014366, EY017373, EY12974, EY14599.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 1994. doi:https://doi.org/
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      C. Lan, Y. Peng, H. Shao, Y. Cui, H. J. Kaplan, D. Sun; Major Role of T Cells in the Induction of Il-17+ Uveitogenic T Cells in Mouse Eau. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1994. doi: https://doi.org/.

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

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Abstract

Purpose: : We have previously demonstrated that both IFN-γ+ and IL-17+ IRBP-specific T cells play a major role in the pathogenesis of experimental autoimmune uveitis (EAU). Recently, we have found that a major portion of the IL-17 producing T cells isolated from B6 mice immunized with an uveitogenic IRBP peptide (IRBP1-20) expressed γΔ TCR. We therefore determined the role of γΔ T cells in the pathogenesis of EAU.

Methods: : γΔ T cells were positively sorted out using magnetic sorter from the IRBP1-20-primed splenic T cells 5 days after the in vitro stimulation with immunizing antigen. To depletion γΔ T cells in vivo, naive mice were treated either with antibody specific for γΔ TCR (GL3). For intracellularly staining of IL-17+ T cells, T cells subjected for analysis were stimulated in vitro with 50 ng/ml of PMA, 1 µg/ml of ionomycin, and 1µg/ml of brefeldin A for 4 h, then were washed, fixed, permeabilized overnight with Cytofix/Cytoperm buffer, and stained with antibodies against IFN-γ and IL-17 and analyzed on a FACScalibur flow cytometer.

Results: : Numbers of peripherally circulating γΔ T cells increased rapidly in B6 mice immunized with the uveitogenic peptide IRBP1-20, which expanded vigorously during subsequent in vitro culture. Highly purified αβ and γΔ T cells produced low levels of IL-17 after exposure to the immunizing antigen in vitro; however, responder T cells mixed with αβ and γΔ T cells at varying ratios produced greatly increased amount of IL-17. Depletion of γΔ T cells of the EAU-prone B10RIII mice drastically decreased the disease severity as well as the IL-17 production of the in vivo primed IRBP-specific T cells.

Conclusions: : Our results show that γΔ T cells are important immune components in the generation and activation of IL-17-producing autoreactive T cells as well as in the pathogenesis of EAU.

Keywords: immunomodulation/immunoregulation • uveitis-clinical/animal model • inflammation 
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