June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Deficiency in Nod2 is associated with dysregulation of Th17-related responses in the eye
Author Affiliations & Notes
  • Ellen J Lee
    Mail code: R&D 14, VA Portland Health Care System, Portland, Oregon, United States
    Molecular Microbiology & Immunology, Oregon Health & Science University, Portland, Oregon, United States
  • Paige Snow
    Mail code: R&D 14, VA Portland Health Care System, Portland, Oregon, United States
  • Joao M Furtado
    Ribeirao Preto Medical School, University of Sao Paulo, Ribeirao Preto, Brazil
  • Ruth Napier
    Molecular Microbiology & Immunology, Oregon Health & Science University, Portland, Oregon, United States
    Mail code: R&D 14, VA Portland Health Care System, Portland, Oregon, United States
  • Emily E Vance
    Molecular Microbiology & Immunology, Oregon Health & Science University, Portland, Oregon, United States
    Mail code: R&D 14, VA Portland Health Care System, Portland, Oregon, United States
  • Phyllis Silver
    Laboratory of Immunology, National Eye Institute, Bethesda, Maryland, United States
  • Justine Smith
    School of Medicine, Flinders University, Adelaide, South Australia, Australia
  • Rachel R Caspi
    Laboratory of Immunology, National Eye Institute, Bethesda, Maryland, United States
  • Holly Lallman Rosenzweig
    Mail code: R&D 14, VA Portland Health Care System, Portland, Oregon, United States
    Molecular Microbiology & Immunology, Oregon Health & Science University, Portland, Oregon, United States
  • Footnotes
    Commercial Relationships   Ellen Lee, None; Paige Snow, None; Joao Furtado, None; Ruth Napier, None; Emily Vance, None; Phyllis Silver, None; Justine Smith, None; Rachel Caspi, None; Holly Rosenzweig, None
  • Footnotes
    Support  NIH/NEI (grant EY025250), the Department of Veterans Affairs Biomedical Laboratory (I01 BX002180; IK2 BX001295); NEI intramural support (Project # EY00184), and ARC (FT130101648)
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 539. doi:
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    • Get Citation

      Ellen J Lee, Paige Snow, Joao M Furtado, Ruth Napier, Emily E Vance, Phyllis Silver, Justine Smith, Rachel R Caspi, Holly Lallman Rosenzweig; Deficiency in Nod2 is associated with dysregulation of Th17-related responses in the eye. Invest. Ophthalmol. Vis. Sci. 2017;58(8):539.

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

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Abstract

Purpose : Mutation in NOD2 results in Blau syndrome, which is characterized by granulomatous uveitis in association with dermatitis and arthritis. NOD2 belongs to a family of innate immune receptors that function in host defense, but we recently identified a novel suppressive role for Nod2 in experimental autoimmune uveitis (EAU), a T cell-dependent model of uveitis. Nod2 deficiency resulted in greater expansion, but similar proportion, of CD4+ T cells in eyes. Here we sought to study the cellular mechanisms behind modulation of EAU by Nod2.

Methods : Mice deficient in Nod2 or congenic wild type (WT) C57BL/6J controls were immunized for EAU with a mixture of interphotoreceptor retinoid-binding protein (IRBP) and IRBP1-20 peptide. Flow cytometric analysis of CD4+ T cells isolated from whole inflamed eyes harvested d21 post-immunization was used to evaluate Th1 (IFNγ+) and Th17 (IL-17+) effector subsets. Single cell suspensions isolated from inflamed eyes were re-stimulated in vitro with IRBP1-20 peptide to assess antigen-recall cytokine responses by flow cytometry after intracellular cytokine staining for IL-17A, TNF, IFNγ, GM-CSF, and IL-22. Relative contributions of IL-17 and IFNγ to disease in vivo were evaluated by fundus imaging and histopathology following antibody neutralization (100 μg/dose), on days -1,0,2,5,9, and 13 relative to immunization.

Results : Nod2 deficiency resulted in increased percentages of ocular CD4+ T cells that produced IL-17 (6.1% vs. 1.52% in WT) or IFNγ (9.38% vs. 1.57% in WT). Antigen-recall studies of purified ocular cells revealed a dramatic increase in the proportion of IL-17+CD4+ cells in Nod2 KO compared to WT mice (12.3% vs. 3.5%). Further evaluation of IRBP-stimulated cytokine production of IL-17+CD4+ cells revealed enhanced production of all cytokines tested, including IFNγ (i.e. IL-17+IFNγ+ cells). Blockade of IL-17 in vivo significantly diminished EAU severity for both genotypes, but to a greater extent in Nod2 KO mice. In contrast, blockade of IFNγ significantly worsened EAU severity in WT mice, but did not alter disease severity in Nod2 KO mice.

Conclusions : These data suggest that the exacerbated uveitis accompanying Nod2 deficiency is mediated primarily through dysregulation of the Th17-related rather than Th1-related response.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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