Detection of autoreactive T cells in type 1 diabetes using coded autoantigens and an immunoglobulin-free cytokine ELISPOT assay: report from the fourth immunology of diabetes society T cell workshop

Ann N Y Acad Sci. 2004 Dec:1037:10-5. doi: 10.1196/annals.1337.002.

Abstract

The "gold standard" for evaluation of immunoassays is blinded testing, using coded samples and antigens. Although assays for autoreactive T cells are no exception to this rule, it is nonetheless rarely applied in this context. To facilitate such investigations, we coded a panel of 10 peptides representing T cell epitopes reported to be of relevance to islet autoimmunity. These were deployed in a novel cytokine ELISPOT assay, in which the use of immunoglobulin-free medium reduces background reactivity and thus potentially enhances the specificity and sensitivity of detection of autoreactive T cells. Significant IFN-gamma production against GAD65 (554-575), insulin (B9-23), and IA-2 (709-736) peptides were observed in type 1 diabetic patients, whereas no significant changes from background were detected in healthy controls. These results confirm the utility of the blinded performance of T cell assays as the most robust platform for assessing technologies to T cell autoreactivity.

Publication types

  • Comparative Study

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Autoantigens / immunology*
  • Case-Control Studies
  • Cells, Cultured
  • Coculture Techniques
  • Cytokines / blood*
  • Cytokines / metabolism
  • Diabetes Mellitus, Type 1 / immunology
  • Diabetes Mellitus, Type 1 / metabolism*
  • Feasibility Studies
  • Female
  • Glutamate Decarboxylase / pharmacology
  • Humans
  • Immunoassay / methods*
  • Insulin / pharmacology
  • Interferon-gamma / biosynthesis
  • Interferon-gamma / immunology
  • Japan
  • Male
  • Membrane Proteins / immunology
  • Middle Aged
  • Peptide Fragments / pharmacology
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1
  • Protein Tyrosine Phosphatases / immunology
  • Receptor-Like Protein Tyrosine Phosphatases, Class 8
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sensitivity and Specificity
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology*

Substances

  • Autoantigens
  • Cytokines
  • Insulin
  • Membrane Proteins
  • Peptide Fragments
  • Recombinant Proteins
  • Interferon-gamma
  • PTPRN protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1
  • Protein Tyrosine Phosphatases
  • Receptor-Like Protein Tyrosine Phosphatases, Class 8
  • Glutamate Decarboxylase