Absence of autoreactive CD4+ T-cells targeting HLA-DQA1*01:02/DQB1*06:02 restricted hypocretin/orexin epitopes in narcolepsy type 1 when detected by EliSpot

J Neuroimmunol. 2017 Aug 15:309:7-11. doi: 10.1016/j.jneuroim.2017.05.001. Epub 2017 May 4.

Abstract

Narcolepsy type 1, a neurological sleep disorder strongly associated with Human Leukocyte Antigen (HLA-)DQB1*06:02, is caused by the loss of hypothalamic neurons producing the wake-promoting neuropeptide hypocretin (hcrt, also known as orexin). This loss is believed to be caused by an autoimmune reaction. To test whether hcrt itself could be a possible target in the autoimmune attack, CD4+ T-cell reactivity towards six different 15-mer peptides from prepro-hypocretin with high predicted affinity to the DQA1*01:02/DQB1*06:02 MHC class II dimer was tested using EliSpot in a cohort of 22 narcolepsy patients with low CSF hcrt levels, and 23 DQB1*06:02 positive healthy controls. Our ELISpot assay had a detection limit of 1:10,000 cells. We present data showing that autoreactive CD4+ T-cells targeting epitopes from the hcrt precursor in the context of MHC-DQA1*01:02/DQB1*06:02 are either not present or present in a frequency is <1:10,000 among peripheral CD4+ T-cells from narcolepsy type 1 patients.

Keywords: CD4(+) T-cells; ELISpot; HLA-DQ; Hypocretin; Narcolepsy; Orexin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Amino Acid Sequence
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • Child
  • Cohort Studies
  • Enzyme-Linked Immunospot Assay / methods*
  • Epitopes / immunology
  • Epitopes / metabolism*
  • Female
  • HLA-DQ alpha-Chains / blood*
  • HLA-DQ alpha-Chains / immunology
  • HLA-DQ beta-Chains / blood*
  • HLA-DQ beta-Chains / immunology
  • Humans
  • Male
  • Middle Aged
  • Narcolepsy / blood*
  • Narcolepsy / diagnosis
  • Narcolepsy / immunology
  • Orexins / blood*
  • Orexins / immunology
  • Young Adult

Substances

  • Epitopes
  • HCRT protein, human
  • HLA-DQ alpha-Chains
  • HLA-DQ beta-Chains
  • HLA-DQA1 antigen
  • HLA-DQB1 antigen
  • Orexins

Supplementary concepts

  • Narcolepsy 1