June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
ERAP1-mediated immunogenicity and immune-phenotypes in HLA-B51+ Behcet’s and Behcet’s uveitis point to pathogenic CD8 T cell effector responses
Author Affiliations & Notes
  • Johannes Nowatzky
    Medicine, Division of Rheumatology, New York University School of Medicine, New York, New York, United States
  • Ann Cavers
    Medicine, Division of Rheumatology, New York University School of Medicine, New York, New York, United States
  • Yesim Özgüler
    Istanbul University Cerrahpasa Medical School, Turkey
  • Arshed Fahad Al-Obeidi
    Medicine, Division of Rheumatology, New York University School of Medicine, New York, New York, United States
  • Berna Yurttas
    Istanbul University Cerrahpasa Medical School, Turkey
  • Hua Zhong
    Population Health - Division of Statistics, NYU Medical Center, New York, United States
  • Yuhe Xia
    Population Health - Division of Statistics, NYU Medical Center, New York, United States
  • Beatrix Ueberheide
    Proteomics Laboratory, NYU Medical Center - Division of Advanced Research and Technology, New York, United States
  • Gülen Hatemi
    Istanbul University Cerrahpasa Medical School, Turkey
  • Matthias Kugler
    Division of Pulmonary Medicine and Critical Care, New York University School of Medicine, New York, New York, United States
  • Olivier Manches
    Insitute for Advanced Biosciences, Inserm U 1209, France
  • Footnotes
    Commercial Relationships   Johannes Nowatzky, None; Ann Cavers, None; Yesim Özgüler, None; Arshed Al-Obeidi, None; Berna Yurttas, None; Hua Zhong, None; Yuhe Xia, None; Beatrix Ueberheide, None; Gülen Hatemi, None; Matthias Kugler, None; Olivier Manches, None
  • Footnotes
    Support  NIH-NEI K08EY025324 (Nowatzky)
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 4300. doi:
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      Johannes Nowatzky, Ann Cavers, Yesim Özgüler, Arshed Fahad Al-Obeidi, Berna Yurttas, Hua Zhong, Yuhe Xia, Beatrix Ueberheide, Gülen Hatemi, Matthias Kugler, Olivier Manches; ERAP1-mediated immunogenicity and immune-phenotypes in HLA-B51+ Behcet’s and Behcet’s uveitis point to pathogenic CD8 T cell effector responses. Invest. Ophthalmol. Vis. Sci. 2020;61(7):4300.

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

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Abstract

Purpose : HLA-B51 is a definite risk factor for Behcet’s uveitis (BU) and Behcet’s disease (BD). A coding variant of ERAP1, Hap10 – with low peptide-trimming activity – vastly potentiates this risk, but is mechanistically unclear. We hypothesized that low or absent ERAP1 activity alters CD8 T cell immunogenicity through changes in the HLA-B51 peptidome and shapes the CD8 T cell immune response in affected subjects.

Methods : We generated HLA-B51+ ERAP1 KO LCL clones using CRISPR-Cas9, performed mass spectrometry of the immunoprecipitated MHC-class I peptidome with subsequent computational deconvolution for HLA-B51-binding peptides. We then assessed single cell (ICS), bulk (ELISA) and proliferative (CFSE) CD8 effector (IFNg) T cell responses through stimulation of allogeneic donor cells with WT vs KO LCL and determined ERAP1 haplotypes in 49 untreated Turkish BD/BU subjects and healthy donors (HD) whose PBMC were profiled using 6 multicolor flow cytometry panels.

Results : WT and KO peptidomes differed significantly (p<0.0005 Fisher’s exact test) with a distinctive shift of peptide length frequencies exceeding 9-mer (binding optimum) in the KO vs WT. This held true for computationally deconvoluted HLA-B51 binders. IFNg-secretion from CD8 T cells stimulated with KO LCL was significantly different from WT (ICS, p=0.0068; ELISA, p=0.0067) as was proliferation. Analysis of 133 T, B, NK and monocyte cell populations revealed predominance of CD8 T and NKT cell subsets in HLA-B51+/Hap10+ BD/BU vs HLA-B51+/Hap10- BD/BU and HD, accounting for 80% of all populations reaching significance (p<0.05, Mann-Whitney). Naïve and effector memory CD8 T cell subsets were inversely correlated. Cohen’s effect sizes were large (>0.8) or very large (>1.2).

Conclusions : We show that absence of functional ERAP1 alters human CD8 T cell immunogenicity. This is mediated by an HLA-class I peptidome with propensity for longer peptides above 9mer and suggests loss or de-novo presentation of peptide-HLA-B51 complexes to cognate CD8 TCR. The reciprocal changes in antigen-experienced vs naïve CD8 T cell subsets in affected subjects point to biologic significance of HLA-B51/Hap10 in BD/BU. Collectively, our findings suggest that an altered HLA-B51 peptidome modulates immunogenicity of CD8 effector T cells in ERAP1-Hap10 carriers with BD/BU and identify targets for future drug development.

This is a 2020 ARVO Annual Meeting abstract.

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