title: Validity and reproducibility of measurement of islet autoreactivity by T-cell assays in subjects with early type 1 diabetes.
abstract: OBJECTIVE
  Type 1 diabetes results from an immunemediated destruction of beta-cells, likely to be mediated by T lymphocytes, but the sensitivity, specificity, and other measures of validity of existing assays for islet autoreactive T-cells are not well established. Such assays are vital for monitoring responses to interventions that may modulate disease progression.
  
  
  METHODS
  We studied the ability of cellular assays to discriminate responses in patients with type 1 diabetes and normal control subjects in a randomized blinded study in the U.S. and U.K. We evaluated the reproducibility of these measurements overall and to individual analytes from repeat collections.
  
  
  RESULTS
  Responses in the cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays could differentiate patients from control subjects with odds ratios of 21.7, 3.44, and 3.36, respectively, with sensitivity and specificity as high as 74 and 88%. The class II tetramer and U.S. ELISPOT assays performed less well. Despite the significant association of the responses with type 1 diabetes, the reproducibility of the measured responses, both overall and individual analytes, was relatively low. Positive samples from normal control subjects (i.e., false positives) were generally isolated to single assays.
  
  
  CONCLUSIONS
  The cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays can distinguish responses from patients with type 1 diabetes and healthy control subjects. The limited reproducibility of the measurements overall and of responses to individual analytes may reflect the difficulty in detection of low frequency of antigen-specific T-cells or variability in their appearance in peripheral blood.
authors.affiliation: Yale University, New Haven, Connecticut, USA. kevan.herold@yale.edu
authors.forename: Kevan C
authors.initials: KC
authors.lastname: Herold
authors.forename: Barbara
authors.initials: B
authors.lastname: Brooks-Worrell
authors.forename: Jerry
authors.initials: J
authors.lastname: Palmer
authors.forename: H Michael
authors.initials: HM
authors.lastname: Dosch
authors.forename: Mark
authors.initials: M
authors.lastname: Peakman
authors.forename: Peter
authors.initials: P
authors.lastname: Gottlieb
authors.forename: Helena
authors.initials: H
authors.lastname: Reijonen
authors.forename: Sefina
authors.initials: S
authors.lastname: Arif
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Spain
authors.forename: Clinton
authors.initials: C
authors.lastname: Thompson
authors.forename: John M
authors.initials: JM
authors.lastname: Lachin
mesh_terms: D000293:Adolescent; D000328:Adult; D001323:Autoantibodies / Q000032:analysis; D002648:Child; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D005260:Female; D005822:Genetic Vectors; D006684:HLA-DR Antigens / Q000235:genetics; D006650:Histocompatibility Testing; D006801:Humans; D007111:Immunity, Cellular; D015151:Immunoblotting; D050417:Insulin-Secreting Cells / Q000276:immunology / Q000473:pathology; D007515:Islets of Langerhans / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D012016:Reference Values; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology*; D055815:Young Adult
chemical_list: D001323:Autoantibodies; D006684:HLA-DR Antigens
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